subject:(Adeno associated vector)

The Ohio State University

1. Santangelo, Kelly Susan. Determining the role of interleukin-1β in the Hartley guinea pig model of primary osteoarthritis.

Degree: PhD, Veterinary Biosciences, 2011, The Ohio State University

URL: http://rave.ohiolink.edu/etdc/view?acc_num=osu1299649441

► Osteoarthritis (OA) is a debilitating disease associated with pain and dysfunction that remains an unresolved and widespread source of symptomatic health problems for many individuals,… (more)

▼ Osteoarthritis (OA) is a debilitating disease associated with pain and dysfunction that remains an unresolved and widespread source of symptomatic health problems for many individuals, particularly those 40 years and older. Interleukin-1β (IL-1β) has been cited as a major cytokine involved in OA-related joint degeneration. Recognized as one of the most important mediators of inflammation and host response to infection, increased production of this cytokine has been linked to a wide variety of autoimmune conditions and autoinflammatory disorders. Characterization of its role in primary OA, however, remains elusive and potentially contradictory. Thus, the molecular interactions to explain the relationship between IL-1β and maintenance of healthy articular cartilage have been proposed but are not yet definitively established, and characterization of the function of IL-1β in a spontaneous, in vivo model remains elusive. As such, a primary aim of this study was to provide a comprehensive analysis of the temporal expression and tissue distribution of IL-1β using immunohistochemistry (IHC) throughout initiation and progression of OA in a naturally-occurring animal model. We subsequently elucidated that OA-prone Hartley guinea pig did not demonstrate reduced IL-1β in weight-bearing articular cartilage, synovium, meniscus, or subchondral bone during achievement of adult maturity as in the control guinea pig strain, and that this aberrant expression may correlate to early incidence of OA. As this temporal study provided evidence that IL-1β is a biomarker relevant to the development and progression of OA, and we then performed in vitro and in vivo studies to block and/or reduce the dysregulation of this cytokine’s expression such that we could provide evidence of its contribution to premature onset of spontaneous OA. Successful reduction of the IL-1β transcript was achieved via RNA interference (RNAi) techniques in vitro using a novel adeno-associated viral vector serotype 5 (AAV5) vector containing a targeting knockdown sequence validated in our laboratory. Importantly, in vitro decreases in IL-1β influenced the transcript levels of several critical mediators involved in OA pathogenesis in the direction of beneficial disease modification. We then proceeded with in vivo investigations to characterize gene expression changes influenced by either our AAV5 targeting knockdown vector or administration of an adenovirus (Ad) vector encoding the human interleukin-1 receptor antagonist protein (hIRAP). This work demonstrated that a reduction in IL-1β signaling by RNAi or hIRAP via AAV5 and Ad vectors, respectively, was achieved in the Hartley guinea pig model of naturally-occurring OA. Importantly, this manipulation was demonstrated over a 120-day period with no loss of reporter gene expression from either viral vector. Reducing IL-1β significantly decreased expression of three inflammatory mediators and one catabolic agent, while it simultaneously increased the number of a specific anabolic molecule, providing biological evidence that… Advisors/Committee Members: Bertone, Alicia (Advisor).

Subjects/Keywords: Molecular Biology; osteoarthritis; interleukin-1; guinea pig; RNA interference; adeno-associated viral vector; adenoviral vector

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APA (6^th Edition):

Santangelo, K. S. (2011). Determining the role of interleukin-1β in the Hartley guinea pig model of primary osteoarthritis. (Doctoral Dissertation). The Ohio State University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=osu1299649441

Chicago Manual of Style (16^th Edition):

Santangelo, Kelly Susan. “Determining the role of interleukin-1β in the Hartley guinea pig model of primary osteoarthritis.” 2011. Doctoral Dissertation, The Ohio State University. Accessed January 19, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=osu1299649441.

MLA Handbook (7^th Edition):

Santangelo, Kelly Susan. “Determining the role of interleukin-1β in the Hartley guinea pig model of primary osteoarthritis.” 2011. Web. 19 Jan 2021.

Vancouver:

Santangelo KS. Determining the role of interleukin-1β in the Hartley guinea pig model of primary osteoarthritis. [Internet] [Doctoral dissertation]. The Ohio State University; 2011. [cited 2021 Jan 19]. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1299649441.

Council of Science Editors:

Santangelo KS. Determining the role of interleukin-1β in the Hartley guinea pig model of primary osteoarthritis. [Doctoral Dissertation]. The Ohio State University; 2011. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1299649441

University of Georgia

2. Estevez, Carlos. Molecular characterization of the avian adeno-associated virus and its use for the development of a viral vector system.

Degree: 2014, University of Georgia

URL: http://hdl.handle.net/10724/21488

► The usefulness of viral-vectored delivery of genetic information to cells and tissues, both in vivo and in vitro, has been well documented. Advances in recombinant… (more)

Subjects/Keywords: Avian adeno-associated virus; Recombinant DNA; Virus vector; Gene delivery; Gene expression.

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APA (6^th Edition):

Estevez, C. (2014). Molecular characterization of the avian adeno-associated virus and its use for the development of a viral vector system. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/21488

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Estevez, Carlos. “Molecular characterization of the avian adeno-associated virus and its use for the development of a viral vector system.” 2014. Thesis, University of Georgia. Accessed January 19, 2021. http://hdl.handle.net/10724/21488.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Estevez, Carlos. “Molecular characterization of the avian adeno-associated virus and its use for the development of a viral vector system.” 2014. Web. 19 Jan 2021.

Vancouver:

Estevez C. Molecular characterization of the avian adeno-associated virus and its use for the development of a viral vector system. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Jan 19]. Available from: http://hdl.handle.net/10724/21488.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Estevez C. Molecular characterization of the avian adeno-associated virus and its use for the development of a viral vector system. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/21488

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

The Ohio State University

3. Cataldi, Marcela Patricia. Diverse Effects of DNA Repair Pathways on the Outcome of Recombinant Adeno-Associated Virus (rAAV) Vector Gene Delivery.

Degree: PhD, Molecular, Cellular and Developmental Biology, 2011, The Ohio State University

URL: http://rave.ohiolink.edu/etdc/view?acc_num=osu1303842573

► Because of Adeno-Associated Virus (AAV) nonpathogenicity and ability to promote sustained, long-term transgene expression in a wide variety of tissues including the brain, liver, muscle,… (more)

▼ Because of Adeno-Associated Virus (AAV) nonpathogenicity and ability to promote sustained, long-term transgene expression in a wide variety of tissues including the brain, liver, muscle, retina, and vasculature, several recombinant AAV (rAAV) serotypes are emerging as attractive vectors for gene therapy. The linear DNA genomes of AAV are acted upon by multiple DNA repair and recombination pathways once released into the host nucleus. The palindromic terminal repeats (TR) on both ends of the AAV genome, which form DNA hairpin (HP) structures, serve as substrates for intramolecular and intermolecular recombination, leading to circularization and concatemer formation, respectively. Infrequently, AAV TRs can also recombine with the host genome, leading to chromosomal integration. However, the mechanism of recombination between AAV TRs is still unclear, and with respect to integration, recombination raises concern for genotoxicity.I have compared the fates of single-stranded rAAV (ssAAV) and self-complementary (scAAV) genomes in cell lines deficient in each of three signaling factors, ATM, ATR, and DNA-PKcs, which orchestrate major DNA double-strand break (DSB) repair pathways. The results suggest that there are qualitative and quantitative differences between both vectors in terms of their interactions with different host DNA repair and recombination pathways. A striking and unexpected observation from these experiments was that in cells deficient in ATM, transduction scored by GFP expression was increased relative to wild-type (WT) cells for both rAAV vectors. The augmented transduction was not reflected in Southern blots of nuclear vector DNA, suggesting that interactions with ATM lead to silencing in normal cells. The additional functional genomes in ATM-/- cells remained linear, and the number of circularized genomes was not affected by the mutation, consistent with compartmentalization of genomes into different DNA repair pathways. To investigate whether the TR HP has any specific effect on recombination, I compared DNA substrates containing the TR sequences in an open DNA duplex conformation or in a covalently closed hairpin conformation, or containing blunt DNA ends with no TR sequence. These DNA substrates were transfected into either WT or DNA-repair deficient cells. DNA molecules with the TR sequences constrained in the hairpin conformation at one or both ends were subject to a loss of gene expression, which was partially relieved in ATM-/- cells. This ATM-mediated effect was not due to a loss of DNA, suggesting that a single AAV TR in the hairpin conformation in cis is necessary and sufficient to mediate the ATM-dependent silencing of gene expression from rAAV genomes. The results also indicate that DNA substrates with AAV TR in a hairpin conformation circularize and integrate with the same efficiency as other forms of DNA ends in WT cells. Therefore, the specificity of the recognition of the HP structure of rAAV vector genomes by DNA repair/recombination pathways leads to loss of functional genomes rather… Advisors/Committee Members: McCarty, Douglas (Advisor).

Subjects/Keywords: Molecular Biology; Virology; adeno-associated virus; gene therapy; DNA recombination; rAAV vector; DNA viral vectors

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Cataldi, M. P. (2011). Diverse Effects of DNA Repair Pathways on the Outcome of Recombinant Adeno-Associated Virus (rAAV) Vector Gene Delivery. (Doctoral Dissertation). The Ohio State University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=osu1303842573

Chicago Manual of Style (16^th Edition):

Cataldi, Marcela Patricia. “Diverse Effects of DNA Repair Pathways on the Outcome of Recombinant Adeno-Associated Virus (rAAV) Vector Gene Delivery.” 2011. Doctoral Dissertation, The Ohio State University. Accessed January 19, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=osu1303842573.

MLA Handbook (7^th Edition):

Cataldi, Marcela Patricia. “Diverse Effects of DNA Repair Pathways on the Outcome of Recombinant Adeno-Associated Virus (rAAV) Vector Gene Delivery.” 2011. Web. 19 Jan 2021.

Vancouver:

Cataldi MP. Diverse Effects of DNA Repair Pathways on the Outcome of Recombinant Adeno-Associated Virus (rAAV) Vector Gene Delivery. [Internet] [Doctoral dissertation]. The Ohio State University; 2011. [cited 2021 Jan 19]. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1303842573.

Council of Science Editors:

Cataldi MP. Diverse Effects of DNA Repair Pathways on the Outcome of Recombinant Adeno-Associated Virus (rAAV) Vector Gene Delivery. [Doctoral Dissertation]. The Ohio State University; 2011. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1303842573

University of Iowa

4. Steines, Benjamin Richard. Investigation and application of novel adeno-associated viral vectors for cystic fibrosis gene therapy.

Degree: PhD, Molecular and Cellular Biology, 2015, University of Iowa

URL: https://ir.uiowa.edu/etd/1763

► Cystic Fibrosis (CF) is a lethal autosomal recessive genetic disorder caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR transports… (more)

▼ Cystic Fibrosis (CF) is a lethal autosomal recessive genetic disorder caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR transports anions at the apical surface of epithelial membranes and functions in many areas of the body. However in CF, loss of CFTR function in the lungs is the major source of morbidity and mortality. Replacing the defective CFTR in the lungs through gene therapy has the potential to cure the disease. Recombinant adeno-associated virus (AAV) is an effective gene transfer vector and has been used extensively to deliver genes to cells in culture. A number of clinical trials using AAV have been attempted for a variety of diseases, including CF, albeit with limited success. Poor vector transduction efficiency prevents effective gene therapy. We have previously used a technique to greatly increase the transduction efficiency of AAV in human lung tissues by selecting from a library of AAVs using a directed evolution technique. However, this evolution was performed in cultured cells and did not fully represent the in vivo environment in which the AAV would be used. In 2008, a CF pig model was developed to develop a further understanding of the mechanisms of CF and CFTR function. We hypothesized that we could use directed evolution to select for a vector in vivo using the pig, allowing gene therapy studies to be conducted in a physiologically relevant model of CF. We selected a novel AAV variant, called AAV2H22, which is closely related to AAV2 but with greatly increased transduction efficiency in pig airway epithelia. AAV2H22 displayed specific tropism for pig airway epithelia and saturated cell surface receptors, indicating specific binding in those cells. We found that AAV2H22-mediated gene transfer corrected chloride and bicarbonate transport defects both in vitro and in vivo. Importantly, bicarbonate transport was sufficient to normalize pH in the airway surface liquid, resulting in increased bacterial killing likely due to increased activity of antimicrobial peptides. To investigate the mechanics of the increased transduction of AAV2H22, capsid mutants were assayed for transduction efficiency. Two of the five amino acid differences between AAV2 and AAV2H22 lie at the surface and are predicted to alter capsid binding. This is consistent with the results showing specific binding in cultured airway epithelia. This research has important implications for gene therapy and investigations using AAV2H22 will increase our understanding of the biology needed to successfully treat CF. Advisors/Committee Members: Zabner, Joseph (supervisor).

Subjects/Keywords: publicabstract; AAV vector; Adeno-associated virus; CF pig model; Cystic fibrosis; Directed evolution; Gene therapy; Cell Biology

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Steines, B. R. (2015). Investigation and application of novel adeno-associated viral vectors for cystic fibrosis gene therapy. (Doctoral Dissertation). University of Iowa. Retrieved from https://ir.uiowa.edu/etd/1763

Chicago Manual of Style (16^th Edition):

Steines, Benjamin Richard. “Investigation and application of novel adeno-associated viral vectors for cystic fibrosis gene therapy.” 2015. Doctoral Dissertation, University of Iowa. Accessed January 19, 2021. https://ir.uiowa.edu/etd/1763.

MLA Handbook (7^th Edition):

Steines, Benjamin Richard. “Investigation and application of novel adeno-associated viral vectors for cystic fibrosis gene therapy.” 2015. Web. 19 Jan 2021.

Vancouver:

Steines BR. Investigation and application of novel adeno-associated viral vectors for cystic fibrosis gene therapy. [Internet] [Doctoral dissertation]. University of Iowa; 2015. [cited 2021 Jan 19]. Available from: https://ir.uiowa.edu/etd/1763.

Council of Science Editors:

Steines BR. Investigation and application of novel adeno-associated viral vectors for cystic fibrosis gene therapy. [Doctoral Dissertation]. University of Iowa; 2015. Available from: https://ir.uiowa.edu/etd/1763

University of Pennsylvania

5. Giles, April. Immunological And Biochemical Evaluation Of The Aav Capsid To Advance Next-Generation Gene Therapy Vector Design.

Degree: 2018, University of Pennsylvania

URL: https://repository.upenn.edu/edissertations/2702

► Gene therapy utilizing adeno-associated viral (AAV) vectors has experienced much success in the clinic recently, culminating in the approval of Luxturna, the first AAV gene… (more)

▼ Gene therapy utilizing adeno-associated viral (AAV) vectors has experienced much success in the clinic recently, culminating in the approval of Luxturna, the first AAV gene replacement product approved by the FDA. However, many significant obstacles remain to the translation of AAV vectors into widely available therapeutics, including immune responses to the capsid as well as a general lack of biochemical characterization of the AAV capsid itself. Humoral immunity, generated in response to viral infection or vector delivery, can reduce therapeutic efficacy due to AAV neutralization by neutralizing antibodies (NAbs). To study these interactions and to inform the design of novel capsids able to evade NAbs, we identified the novel and highly neutralizing antibody PAV9.1 from a panel of hybridoma clones and mapped its epitope on AAV9 by cryo-EM. Mutagenesis efforts within this epitope established the minimal changes required to confer evasion of both PAV9.1 binding and neutralization, but this evasion was not found to extend to polyclonal, NAb positive samples from a number of sources, reflecting the complexity of the NAb repertoire against AAV. To enable more complete study of this repertoire in a more therapeutically-relevant fashion, we also established a pipeline for the unbiased cloning of novel α-AAV antibodies from single memory B cells isolated from seropositive individuals. Using this approach, we were able to isolate a panel of mAbs found to recognize AAV capsid, validating the use of this method for further study of the polyclonal AAV Ab repertoire. Finally, to come to a better understanding of the biochemical properties of the AAV capsid with the potential to influence critical aspects of vector performance, including cellular transduction and interactions with the immune system, we evaluated AAV by mass spectrometry for the presence of post-translational modifications. We established that there is extensive deamidation of asparagine residues and through genetic deamidation mutagenesis determined that these spontaneous events are a source of potentially undesirable heterogeneity that modulates capsid protein charge, assembly, and in vitro and in vivo transduction. These studies highlight novel findings that contribute to the greater understanding of AAV biology and will inform future efforts to develop next-generation vectors.

Subjects/Keywords: Adeno-associated virus; Gene therapy; Viral vector; Allergy and Immunology; Immunology and Infectious Disease; Medical Immunology; Molecular Biology; Virology

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Giles, A. (2018). Immunological And Biochemical Evaluation Of The Aav Capsid To Advance Next-Generation Gene Therapy Vector Design. (Thesis). University of Pennsylvania. Retrieved from https://repository.upenn.edu/edissertations/2702

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Giles, April. “Immunological And Biochemical Evaluation Of The Aav Capsid To Advance Next-Generation Gene Therapy Vector Design.” 2018. Thesis, University of Pennsylvania. Accessed January 19, 2021. https://repository.upenn.edu/edissertations/2702.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Giles, April. “Immunological And Biochemical Evaluation Of The Aav Capsid To Advance Next-Generation Gene Therapy Vector Design.” 2018. Web. 19 Jan 2021.

Vancouver:

Giles A. Immunological And Biochemical Evaluation Of The Aav Capsid To Advance Next-Generation Gene Therapy Vector Design. [Internet] [Thesis]. University of Pennsylvania; 2018. [cited 2021 Jan 19]. Available from: https://repository.upenn.edu/edissertations/2702.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Giles A. Immunological And Biochemical Evaluation Of The Aav Capsid To Advance Next-Generation Gene Therapy Vector Design. [Thesis]. University of Pennsylvania; 2018. Available from: https://repository.upenn.edu/edissertations/2702

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

6. Rossi, Axel. Intracellular fate of AAV particles in human Dendritic Cell and impact on Gene Transfer : Devenir intracellulaire des vecteurs AAV dans les cellules dendritiques humaines et conséquences sur le transfert de gène.

Degree: Docteur es, Sciences de la Vie, 2016, Lyon

URL: http://www.theses.fr/2016LYSEN028

►

Les vecteurs viraux dérivés du virus adéno-associé (AAV) apparaissent depuis deux décennies, comme des outils efficaces pour le transfert de gène in vivo. Cependant, malgré… (more)

▼

Les vecteurs viraux dérivés du virus adéno-associé (AAV) apparaissent depuis deux décennies, comme des outils efficaces pour le transfert de gène in vivo. Cependant, malgré une faible immunogénicité et une absence de toxicité in vivo, leur optimisation requiert encore un effort important vers une meilleure compréhension de leur biologie et, en particulier, de leur interaction avec le système immunitaire. Au cours de ce travail de thèse, nous avons utilisé une méthode de sélection dirigée in vitro dans le but d’obtenir un variant de capside capable de transduire efficacement un type cellulaire non-permissif aux vecteurs AAV : les cellules dendritiques (DC). En effet, ces cellules jouent un rôle primordial dans l’établissement de la réponse immunitaire et, par conséquent, dans la persistance de l’expression du transgène in vivo. Cette technologie, très répandue dans la communauté AAV, a permis de sélectionner un variant de capside aux propriétés très intéressantes. La mutation sélectionnée, caractérisée in vitro comme induisant une instabilité de la capside, a permis d’identifier et de surmonter un point de blocage majeur dans le processus de transduction des DC par les vecteurs AAV consistant dans l’étape de décapsidation du génome du vecteur dans le noyau cellulaire. De manière intéressante, le variant obtenu exhibe un avantage en terme de transduction non seulement dans les DC mais aussi dans différents modèles de cellules primaires humaines (e.g. HUVEC) ou animales (OBC), peu ou pas permissive à l’AAV. De plus, des expériences de transfert de gène in vivo réalisées dans un modèle murin, indiquent que le variant sélectionné conduit à une meilleure expression du transgène, possiblement due à la mise en place d’un processus de tolérisation. Les propriétés remarquables de ce variant de capside, font de lui un candidat intéressant pour des applications médicales.

Vectors derived from the Adeno-associated virus (AAV) have emerged as an efficient system for in vivo gene transfer. However, despite their low immunogenicity and good tolerance in vivo, a better characterization of the host-AAV interaction is required to be able to fully exploit AAV’s potential fora gene therapy or gene vaccination. In this PhD project, we have used an in vitro directed evolution strategy to select an AAV capsid variant able to transduce human dendritic cell (DC), a non-permissive cell type which plays a critical role in the initiation of immune responses and, consequently, on the persistence of the expression of transgene in vivo. This procedure allowed us to identify an AAV variant characterized by a decreased stability of the capsid in vitro. The use of this mutant as a vector to transduce human DC resulted in an improved uncoating of the vector genome in the cell nucleus, thus identifying this step as major barrier toward DC transduction. Interestingly, the selected variant also displayed an increased transduction efficiency not only in DC but also in different primary human and animal cell types, poorly or non-permissive to AAV.…

Advisors/Committee Members: Salvetti, Anna (thesis director), Büning, Hildegard (thesis director).

Subjects/Keywords: Adeno-associated virus; AAV vector; Transport intracellulaire; Ingénierie de capside; Décapsidation; Tolérance; Réponses Immunitaires; Transfert de gène; Adeno-associated virus; AAV vector; Intracellular trafficking; Capsid Engineering; Uncoating; Tolerance; Immune Responses; Gene Transfer

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Rossi, A. (2016). Intracellular fate of AAV particles in human Dendritic Cell and impact on Gene Transfer : Devenir intracellulaire des vecteurs AAV dans les cellules dendritiques humaines et conséquences sur le transfert de gène. (Doctoral Dissertation). Lyon. Retrieved from http://www.theses.fr/2016LYSEN028

Chicago Manual of Style (16^th Edition):

Rossi, Axel. “Intracellular fate of AAV particles in human Dendritic Cell and impact on Gene Transfer : Devenir intracellulaire des vecteurs AAV dans les cellules dendritiques humaines et conséquences sur le transfert de gène.” 2016. Doctoral Dissertation, Lyon. Accessed January 19, 2021. http://www.theses.fr/2016LYSEN028.

MLA Handbook (7^th Edition):

Rossi, Axel. “Intracellular fate of AAV particles in human Dendritic Cell and impact on Gene Transfer : Devenir intracellulaire des vecteurs AAV dans les cellules dendritiques humaines et conséquences sur le transfert de gène.” 2016. Web. 19 Jan 2021.

Vancouver:

Rossi A. Intracellular fate of AAV particles in human Dendritic Cell and impact on Gene Transfer : Devenir intracellulaire des vecteurs AAV dans les cellules dendritiques humaines et conséquences sur le transfert de gène. [Internet] [Doctoral dissertation]. Lyon; 2016. [cited 2021 Jan 19]. Available from: http://www.theses.fr/2016LYSEN028.

Council of Science Editors:

Rossi A. Intracellular fate of AAV particles in human Dendritic Cell and impact on Gene Transfer : Devenir intracellulaire des vecteurs AAV dans les cellules dendritiques humaines et conséquences sur le transfert de gène. [Doctoral Dissertation]. Lyon; 2016. Available from: http://www.theses.fr/2016LYSEN028

Lincoln University

7. Mitchell, Nadia. Longitudinal studies and the development of gene therapy for ovine neuronal ceroid lipofuscinoses.

Degree: 2016, Lincoln University

URL: http://hdl.handle.net/10182/7237

► The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are a group of fatal inherited lysosomal storage diseases characterised by progressive neurodegeneration, cortical atrophy, and blindness. Currently… (more)

▼ The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are a group of fatal inherited lysosomal storage diseases characterised by progressive neurodegeneration, cortical atrophy, and blindness. Currently there are no effective treatments. Naturally occurring animal models exist, including two forms in sheep which are representative of the different NCL protein defects. A lesion in a soluble lysosomal protein (CLN5) causes NCL in Borderdale sheep, whilst South Hampshire sheep have a defective intracellular endoplasmic reticulum membrane-bound protein (CLN6). This subdivision has consequences for the planning of therapies. This thesis compares the progressive neuropathological changes, and examines the efficacy of viral-mediated in vivo gene therapy, in these two NCL sheep models. An immunohistochemical study revealed that, despite very different gene products and subcellular localisations, the pathogenic cascade was remarkably similar for CLN5 and CLN6 affected sheep. Dysregulated glial activation preceded regionally specific neurodegeneration in both disease models and both occurred well before clinical onset. Neuropathological changes were more advanced in the CLN5 model, which correlated with the earlier onset of clinical symptoms in these sheep, but by end-stage disease CLN5 and CLN6 brains were similarly affected. Windows for best therapeutic efficacy were established and these data highlight the translational utility of the sheep brain for testing human gene therapies. Lentiviral vectors have been shown to mediate successful gene delivery to the ovine brain yet the efficacy of adeno-associated viral (AAV) vectors has not been tested in sheep. Stable, predominantly neurotropic, transgene expression was evident one month after intracerebroventricular (ICV) and intraparenchymal (IP) delivery of AAV9 to the normal sheep brain. However with a greater spread and no evidence of vector or procedural neuroinflammation or toxicity, the ICV approach proved most efficacious in sheep. vi Deficiencies in soluble lysosomal proteins are deemed particularly amenable to in vivo gene therapy via the normal lysosomal enzyme trafficking system and the phenomenon of ‘cross-correction’. To test this paradigm in sheep, six pre-clinical CLN5 deficient lambs were treated with combinatorial ICV and IP injections of either lentiviral or AAV9 vectors expressing ovine CLN5. Both vector platforms afforded sustained protection against stereotypical disease in these sheep. Cognitive and neurological function was preserved, whilst longitudinal neuroimaging revealed normalisation of intracranial volume and structural brain integrity. Quality of life was profoundly improved for the treated sheep and, apart from delayed-onset visual deficits, treated sheep well exceeded the typical lifespan of untreated animals. Defects in membrane-bound proteins are generally considered harder therapeutic targets. However the current study indicates that gene therapy is also possible for these NCL forms. Whilst five similarly injected pre-clinical CLN6…

Subjects/Keywords: Batten disease; neuronal ceroid lipofuscinosis; lysosomal storage disorder; animal models; sheep; brain; neurodegeneration; neuroinflammation; neurogenesis; gene therapy; vector; adeno-associated virus; lentivirus; transduction; 1109 Neurosciences; 070709 Veterinary Pathology

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Mitchell, N. (2016). Longitudinal studies and the development of gene therapy for ovine neuronal ceroid lipofuscinoses. (Thesis). Lincoln University. Retrieved from http://hdl.handle.net/10182/7237

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Mitchell, Nadia. “Longitudinal studies and the development of gene therapy for ovine neuronal ceroid lipofuscinoses.” 2016. Thesis, Lincoln University. Accessed January 19, 2021. http://hdl.handle.net/10182/7237.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Mitchell, Nadia. “Longitudinal studies and the development of gene therapy for ovine neuronal ceroid lipofuscinoses.” 2016. Web. 19 Jan 2021.

Vancouver:

Mitchell N. Longitudinal studies and the development of gene therapy for ovine neuronal ceroid lipofuscinoses. [Internet] [Thesis]. Lincoln University; 2016. [cited 2021 Jan 19]. Available from: http://hdl.handle.net/10182/7237.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Mitchell N. Longitudinal studies and the development of gene therapy for ovine neuronal ceroid lipofuscinoses. [Thesis]. Lincoln University; 2016. Available from: http://hdl.handle.net/10182/7237

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Otago

8. Cheong, Isaiah. Gene therapy for Alzheimer's disease: Characterising lentivirus and adeno-associated virus spread from the adult mouse hippocampus .

Degree: University of Otago

URL: http://hdl.handle.net/10523/6288

► Alzheimer’s Disease is a neurodegenerative condition with progressively worsening memory and cognitive function, which ultimately results in death. The two major neuropathological hallmarks of AD… (more)

▼ Alzheimer’s Disease is a neurodegenerative condition with progressively worsening memory and cognitive function, which ultimately results in death. The two major neuropathological hallmarks of AD are extracellular amyloid-beta (Aβ) plaques and intracellular neurofibrillary tau tangles (NFTs). Other pathways and neuronal mechanisms are also likely to be affected as AD is a complex disease resulting from anomalies in components of different pathways. Currently, apart from a few FDA approved drugs which serve to delay symptom progression through slowing neurotransmitter breakdown (Cholinesterase inhibitors) and cell damage (NMDA-receptor antagonist), no effective treatment of the underlying causes of AD is available. One of the hypothesised underlying cause of AD is the formation of extracellular Aβ plaques. Current work in the Neural Development and Disease (NDD) lab have researched viral-mediated gene therapy as a means to introduce a neuroprotective secreted amyloid precursor protein α (sAPPα) gene intracranially. This project’s primary aim was optimising and comparing the spread between lentiviral (LV) and adeno-associated viral (AAV) vectors; along with the effects of systemically delivering an osmotic agent, mannitol. Viral spread would be determined by the use of a GFP reporter gene, where expression indicated transduction and extent of viral spread. This project has showed a significant improvement in transduction efficiency when using an AAV vector to deliver a reporter gene. It efficiently achieved widespread transduction of the CNS from a unilateral injection at the hippocampus. However, while LV had a limited spread in comparison to AAV9, it would be useful for targeted delivery of treatment. Mannitol did not produce any significant effect on vector spread. The secondary aim was to create a plasmid to visualise secretion of sAPPα from virally transduced cells. Once packaged into a lentiviral vector, the plasmid was shown to successfully transduce primary neurons in vitro, but expression of reporter genes were not observed in vivo. Advisors/Committee Members: Hughes, Stephanie (advisor).

Subjects/Keywords: Alzheimer's Disease; Gene therapy; Viral vector; lentivirus; adeno-associated virus; sAPPα; mannitol

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APA (6^th Edition):

Cheong, I. (n.d.). Gene therapy for Alzheimer's disease: Characterising lentivirus and adeno-associated virus spread from the adult mouse hippocampus . (Masters Thesis). University of Otago. Retrieved from http://hdl.handle.net/10523/6288

Note: this citation may be lacking information needed for this citation format:
No year of publication.

Chicago Manual of Style (16^th Edition):

Cheong, Isaiah. “Gene therapy for Alzheimer's disease: Characterising lentivirus and adeno-associated virus spread from the adult mouse hippocampus .” Masters Thesis, University of Otago. Accessed January 19, 2021. http://hdl.handle.net/10523/6288.

Note: this citation may be lacking information needed for this citation format:
No year of publication.

MLA Handbook (7^th Edition):

Cheong, Isaiah. “Gene therapy for Alzheimer's disease: Characterising lentivirus and adeno-associated virus spread from the adult mouse hippocampus .” Web. 19 Jan 2021.

Note: this citation may be lacking information needed for this citation format:
No year of publication.

Vancouver:

Cheong I. Gene therapy for Alzheimer's disease: Characterising lentivirus and adeno-associated virus spread from the adult mouse hippocampus . [Internet] [Masters thesis]. University of Otago; [cited 2021 Jan 19]. Available from: http://hdl.handle.net/10523/6288.

Note: this citation may be lacking information needed for this citation format:
No year of publication.

Council of Science Editors:

Cheong I. Gene therapy for Alzheimer's disease: Characterising lentivirus and adeno-associated virus spread from the adult mouse hippocampus . [Masters Thesis]. University of Otago; Available from: http://hdl.handle.net/10523/6288

Note: this citation may be lacking information needed for this citation format:
No year of publication.

Rice University

9. Brun, Mitchell John. Design of Caspase and MMP-Activatable Adeno-Associated Virus Vectors and Their In Vivo Application.

Degree: PhD, Engineering, 2020, Rice University

URL: http://hdl.handle.net/1911/109218

► Gene therapy is the next evolution in the treatment of diseases, allowing for the correction of genetic mutations, induction of growth to facilitate recovery at… (more)

▼ Gene therapy is the next evolution in the treatment of diseases, allowing for the correction of genetic mutations, induction of growth to facilitate recovery at injury sites, or the delivery of suicide genes to tumor cells. However, the translation of more gene therapies to the clinic has been hindered by the lack of delivery specificity and efficiency. Many viral vectors have been engineered in an attempt to increase transduction efficiency and targeting capabilities. Adeno-associated virus (AAV) has recently become popular as a gene delivery vector because it has the ability to transduce many cell types efficiently, it is non-pathogenic as well as replication deficient, and it is able to be genetically modified. AAV is a promising vector for gene therapy, but its broad tropism can be detrimental if the transgene being delivered is harmful when expressed in non-target tissues. Delivering the transgene of interest to target cells at levels high enough to be effective while maintaining safety by minimizing delivery to off target cells is a prevalent challenge in the field of gene therapy. To address this problem, our lab has developed a protease-activatable vector (provector) platform based on AAV9 that responds to extracellular proteases present at disease sites. This thesis details my work expanding the provector platform to target cysteine-aspartic proteases as well as matrix-metalloproteinases as stimuli for provector activation. These caspase-activatable provectors demonstrate up to 200-fold reduction in transduction ability in the OFF state compared to AAV9, reducing the virus’ ability to transduce healthy tissue. Following proteolysis by caspase-3, the provector shows a 90-fold increase in transduction compared to the OFF state. This provector has also been characterized in vivo, where compared to AAV9 the provector has significantly decreased delivery to off target organs with similar levels of delivery to the injured heart following a myocardial infarction. This work also details characterization of the of the MMP-activatable provector in disease models of acute heart failure, stroke, and chronic heart disease while also explores the therapeutic efficacy of delivering various therapeutic transgenes in murine MI models. To further increase the control of gene delivery with the provector platform, I also detail designs and in vitro testing of provectors requiring two inputs for activation. Preliminary results indicate the vectors perform as designed in vitro which could provide higher specificity of delivery in vivo. Overall, this thesis diversifies the provector platform to target new diseases and increases our understanding of the provector behavior in vitro and in vivo. Advisors/Committee Members: Suh, Junghae (advisor).

Subjects/Keywords: Adeno-associated virus; AAV; gene therapy; targeted gene delivery; viral vector; stimulus-responsive; activatable; de-targeting; capsid engineering; heart disease; apoptosis; protease; cysteine-aspartic proteases; provector

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Brun, M. J. (2020). Design of Caspase and MMP-Activatable Adeno-Associated Virus Vectors and Their In Vivo Application. (Doctoral Dissertation). Rice University. Retrieved from http://hdl.handle.net/1911/109218

Chicago Manual of Style (16^th Edition):

Brun, Mitchell John. “Design of Caspase and MMP-Activatable Adeno-Associated Virus Vectors and Their In Vivo Application.” 2020. Doctoral Dissertation, Rice University. Accessed January 19, 2021. http://hdl.handle.net/1911/109218.

MLA Handbook (7^th Edition):

Brun, Mitchell John. “Design of Caspase and MMP-Activatable Adeno-Associated Virus Vectors and Their In Vivo Application.” 2020. Web. 19 Jan 2021.

Vancouver:

Brun MJ. Design of Caspase and MMP-Activatable Adeno-Associated Virus Vectors and Their In Vivo Application. [Internet] [Doctoral dissertation]. Rice University; 2020. [cited 2021 Jan 19]. Available from: http://hdl.handle.net/1911/109218.

Council of Science Editors:

Brun MJ. Design of Caspase and MMP-Activatable Adeno-Associated Virus Vectors and Their In Vivo Application. [Doctoral Dissertation]. Rice University; 2020. Available from: http://hdl.handle.net/1911/109218

University of Waterloo

10. Cheng, Yu-Lei. Investigation of Sf-9 Cell Metabolism Before and After Baculovirus Infection Using Biovolume: a Case for the Improvement of Adeno-Associated Viral Vector Production.

Degree: 2009, University of Waterloo

URL: http://hdl.handle.net/10012/4540

► Adeno-associated viral (AAV) vectors have been shown to be potential vectors for the treatment of diseases, including protocols using RNA interference (RNAi). AAV vector production… (more)

▼ Adeno-associated viral (AAV) vectors have been shown to be potential vectors for the treatment of diseases, including protocols using RNA interference (RNAi). AAV vector production in insect cells using the baculovirus vector expression system has been a major advance in furthering their use. A major limitation of AAV vector production at high cell densities is a reduction in cell specific yield, which is thought to be caused by nutrient limitations. Nutrient consumption profiles after infection, however, have still not been fully characterized, probably due to the difficulty of characterizing consumption patterns based on increases in cell density, which are minimal after infection. It is known, however, that cells increase in size after infection; therefore, the driving hypothesis of this thesis was that biovolume, or the total volume enclosed by the membrane of viable cells, which accounts for both cell density and cell size, could be used to characterize nutrient consumption patterns both before and after infection. The relationships between nutrient consumption and change in cell density and biovolume were examined by statistical correlation analysis. It was found that in uninfected cultures, no significant correlation differences, using either cell density or biovolume, were observed since cell size remained relatively constant; however, in infected cultures, more than half of the nutrients were found to be better correlated with biovolume than with cell density. When examining the nutrient and metabolite concentration data on a biovolume basis, nutrient consumption remained relatively constant. It is hypothesized that since it has been reported that the rate of cell respiration increases after infection, a more complete oxidation of nutrients occurs to satisfy increased energy needs during infection. By having a basis to base nutrient consumption, we can better assess the needs of the culture. This will allow the development of feeding strategies based on cellular requirements instead of supplying the cultures with generic nutrient cocktails. It is expected that different nutrient mixtures can be used to target different goals such as 1) enhancing cell growth (before infection) and 2) improving the production of recombinant products (after infection). This will not only increase the efficiency of AAV vector production, but will also reduce the cost of production and make the process more economical by eliminating the addition of unnecessary nutrients. Although promising, some limitations of using biovolume still exist. A first limitation is the biovolume measure itself. This measure requires a device that measures cell size, such as a Coulter Counter Multisizer (Beckman-Coulter, Miami, FL, USA), which can be expensive. Capacitance probes can be a more cost effective tool to estimate biovolume; however, the availability of capacitance probes is still not common. A second limitation is the interpretation of the biovolume profiles, which can depend strongly on the fraction of cells in culture that are…

Subjects/Keywords: Sf-9; Metabolism; Baculovirus; Infection; Biovolume; Adeno-Associated Viral Vector; AAV; BEVS

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APA (6^th Edition):

Cheng, Y. (2009). Investigation of Sf-9 Cell Metabolism Before and After Baculovirus Infection Using Biovolume: a Case for the Improvement of Adeno-Associated Viral Vector Production. (Thesis). University of Waterloo. Retrieved from http://hdl.handle.net/10012/4540

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Cheng, Yu-Lei. “Investigation of Sf-9 Cell Metabolism Before and After Baculovirus Infection Using Biovolume: a Case for the Improvement of Adeno-Associated Viral Vector Production.” 2009. Thesis, University of Waterloo. Accessed January 19, 2021. http://hdl.handle.net/10012/4540.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Cheng, Yu-Lei. “Investigation of Sf-9 Cell Metabolism Before and After Baculovirus Infection Using Biovolume: a Case for the Improvement of Adeno-Associated Viral Vector Production.” 2009. Web. 19 Jan 2021.

Vancouver:

Cheng Y. Investigation of Sf-9 Cell Metabolism Before and After Baculovirus Infection Using Biovolume: a Case for the Improvement of Adeno-Associated Viral Vector Production. [Internet] [Thesis]. University of Waterloo; 2009. [cited 2021 Jan 19]. Available from: http://hdl.handle.net/10012/4540.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Cheng Y. Investigation of Sf-9 Cell Metabolism Before and After Baculovirus Infection Using Biovolume: a Case for the Improvement of Adeno-Associated Viral Vector Production. [Thesis]. University of Waterloo; 2009. Available from: http://hdl.handle.net/10012/4540

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

George Mason University

11. Alrashed, Bayan. Targeting HBV cccDNA with Viral Vector Carrying HBV-Specific CRISPR/Cas9 .

Degree: George Mason University

URL: http://hdl.handle.net/1920/10752

► Chronic HBV infection is a worldwide public health concern. It is characterized by the persistence of the hepatitis B surface antigen (HBsAg) in serum for… (more)

▼ Chronic HBV infection is a worldwide public health concern. It is characterized by the persistence of the hepatitis B surface antigen (HBsAg) in serum for more than six months. The persistence of HBsAg is mediated by viral covalently closed circular DNA (cccDNA) in the nuclei of infected cells. Although several genome-editing techniques have demonstrated the possibility to inhibit HBV viral replication, attempts to completely clear nuclear cccDNA remain unsuccessful. Recent studies have suggested the possibility to use the Clustered Regularly Interspaced Short Palindromic Repeats system (CRISPR/Cas9) to suppress HBV infection; however, effective delivery of this tool for in vivo targeting is a major issue. To examine the possibility to utilize the CRISPR/Cas9 system for HBV clearance, we plan to generate a mouse model of chronic HBV infection by using a recombinant Adeno-associated virus that expresses HBV (rAAV-HBV). It has been previously suggested that a rAAV-HBV virus can establish persistent HBV infection in mice. We would like to use this model to test whether viral vectors that carry HBV-specific CRISPR/Cas9 can target HBV cccDNA in vivo. To this end, the recombinant AAV-HBV virus was produced by an AAV helper free expression system in HEK293T cells. As a control, I also produced a rAAV-GFP virus from co-transfection of HEK293T cells. My results show that the recombinant AAV-GFP virus can effectively infect human liver HepG2 cells and express high levels of green fluorescent protein. I also examined the rAAV-HBV virus for expressing HBV genes in HepG2 cells by western blot and quantitative real-time PCR (qPCR). While western blots showed defined bands of HBsAg in HepG2 cells, qPCR of infected HepG2 culture supernatants and cell lysates yielded a low amount of HBV genomes. We therefore demonstrated the possibility to use rAAV-HBV vector to delivery HBV genome into liver cells. However, the efficiency is relatively low, likely resulting from the large size of the HBV genome that may affect the AAV packaging efficiency. Multiple approaches to improve the efficiency of the rAAV-HBV vector are currently being tested. Advisors/Committee Members: Wu, Yuntao (advisor).

Subjects/Keywords: Hepatitus B virus; CCC DNA; Adeno-associated virus; CRISPR/cas9; viral vector; lenti-viral vector

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Alrashed, B. (n.d.). Targeting HBV cccDNA with Viral Vector Carrying HBV-Specific CRISPR/Cas9 . (Thesis). George Mason University. Retrieved from http://hdl.handle.net/1920/10752

Note: this citation may be lacking information needed for this citation format:
No year of publication.
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Alrashed, Bayan. “Targeting HBV cccDNA with Viral Vector Carrying HBV-Specific CRISPR/Cas9 .” Thesis, George Mason University. Accessed January 19, 2021. http://hdl.handle.net/1920/10752.

Note: this citation may be lacking information needed for this citation format:
No year of publication.
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Alrashed, Bayan. “Targeting HBV cccDNA with Viral Vector Carrying HBV-Specific CRISPR/Cas9 .” Web. 19 Jan 2021.

Note: this citation may be lacking information needed for this citation format:
No year of publication.

Vancouver:

Alrashed B. Targeting HBV cccDNA with Viral Vector Carrying HBV-Specific CRISPR/Cas9 . [Internet] [Thesis]. George Mason University; [cited 2021 Jan 19]. Available from: http://hdl.handle.net/1920/10752.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
No year of publication.

Council of Science Editors:

Alrashed B. Targeting HBV cccDNA with Viral Vector Carrying HBV-Specific CRISPR/Cas9 . [Thesis]. George Mason University; Available from: http://hdl.handle.net/1920/10752

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
No year of publication.

12. Dudek, Amanda. A Genome-Wide Knock-Out Screen Identifies Novel Host Cell Entry Factor Requirements for Divergent Adeno-Associated Virus Serotypes.

Degree: PhD, 2019, Harvard University

URL: http://nrs.harvard.edu/urn-3:HUL.InstRepos:41121284

►

Adeno-Associated Virus (AAV) is a non-pathogenic virus that has been harnessed as a vector system for therapeutic gene transfer. Despite decades of research on AAV… (more)

▼

Adeno-Associated Virus (AAV) is a non-pathogenic virus that has been harnessed as a vector system for therapeutic gene transfer. Despite decades of research on AAV as a vector, little is known about the molecular determinants required for cellular entry of different AAV capsid serotypes and how this may lead to unique cell and tissue tropism for these viral vectors. This work presents and characterizes a novel AAV entry pathway used by an evolutionarily related subset of divergent AAVs that is independent of the canonical AAV receptor, AAVR, both in vitro and in vivo. This subset of AAV capsids, comprised of rh32.33 and AAV4, are unable to use AAVR for entry or to bind AAVR in vitro. We further characterize the role of AAVR for AAVR dependent serotypes, suggesting a predominantly post-attachment function for this receptor. Mutagenesis of both capsid and AAVR investigate the role human and animal variation may have in the success and translation of AAV-based gene therapies to the clinic. Additionally, a genome-wide CRISPR/Cas9 based entry screen identifies multiple cellular factors required for entry of AAVR dependent and independent serotypes. First, we describe a minimal alternate receptor complex comprised of NEU1 and CTSA that is uniquely required for entry of AAVR independent serotypes and acts at a post-attachment step in the entry pathway. We also identify and describe a previously uncharacterized protein, required by all AAV serotypes except AAV5. Chimeric capsids suggest both a capsid binding-site as well as an endosomal escape function located within the VP1 unique region of capsid. We therefore present a model in which most AAVs use a canonical entry pathway requiring two conserved entry factors, yet the highly divergent AAV serotypes AAV5, AAV4, and rh32.33 have unique alternate entry factor requirements. Our basic virology studies of AAV aim to both inform currently unanswered questions on the implementation of AAV-based gene therapies in the clinic, as well as allow for better design of AAV-based gene therapies in the future.

Medical Sciences

Advisors/Committee Members: Allen, Todd (advisor), Cepko, Constance (committee member), Wagers, Amy (committee member), Flotte, Terence (committee member).

Subjects/Keywords: Adeno-associated virus; AAV; gene therapy; viral vector; virus; viral entry; CRISPR

…126 V: Implications for adeno-associated viral vector design 127 VI: Concluding remarks… …attractive and safe gene therapy vector. Viral variation of both adenovirus and adeno-associated… …Adeno-associated virus is an attractive viral vector due to its safety profile and has… …was ironically also done using AAV2. VI: The adeno-associated viral vector system Although… …in order to produce a replication defective adeno-associated virus vector [Fig. 1.4.A…

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APA (6^th Edition):

Dudek, A. (2019). A Genome-Wide Knock-Out Screen Identifies Novel Host Cell Entry Factor Requirements for Divergent Adeno-Associated Virus Serotypes. (Doctoral Dissertation). Harvard University. Retrieved from http://nrs.harvard.edu/urn-3:HUL.InstRepos:41121284

Chicago Manual of Style (16^th Edition):

Dudek, Amanda. “A Genome-Wide Knock-Out Screen Identifies Novel Host Cell Entry Factor Requirements for Divergent Adeno-Associated Virus Serotypes.” 2019. Doctoral Dissertation, Harvard University. Accessed January 19, 2021. http://nrs.harvard.edu/urn-3:HUL.InstRepos:41121284.

MLA Handbook (7^th Edition):

Dudek, Amanda. “A Genome-Wide Knock-Out Screen Identifies Novel Host Cell Entry Factor Requirements for Divergent Adeno-Associated Virus Serotypes.” 2019. Web. 19 Jan 2021.

Vancouver:

Dudek A. A Genome-Wide Knock-Out Screen Identifies Novel Host Cell Entry Factor Requirements for Divergent Adeno-Associated Virus Serotypes. [Internet] [Doctoral dissertation]. Harvard University; 2019. [cited 2021 Jan 19]. Available from: http://nrs.harvard.edu/urn-3:HUL.InstRepos:41121284.

Council of Science Editors:

Dudek A. A Genome-Wide Knock-Out Screen Identifies Novel Host Cell Entry Factor Requirements for Divergent Adeno-Associated Virus Serotypes. [Doctoral Dissertation]. Harvard University; 2019. Available from: http://nrs.harvard.edu/urn-3:HUL.InstRepos:41121284

13. Williams, Ryan R. Intrinsic and Extrinsic Factors for the Regeneration of Brainstem Axons after Spinal Cord Injury.

Degree: PhD, Neuroscience (Medicine), 2010, University of Miami

URL: https://scholarlyrepository.miami.edu/oa_dissertations/937

► After traumatic injury, the successful regeneration of axons in the mammalian central nervous system (CNS) that leads to functional recovery will require a combination… (more)

▼ After traumatic injury, the successful regeneration of axons in the mammalian central nervous system (CNS) that leads to functional recovery will require a combination of therapeutic strategies. Following complete transection of the thoracic spinal cord, previous studies found the implantation of a bridge containing Schwann cells (SCs) and Matrigel promotes intraspinal but not supraspinal axon regeneration. Therefore, work in this dissertation sought to elucidate both intrinsic and extrinsic factors that enhance the regeneration of brainstem axons in a SC bridge. Adeno-associated viral (AAV) vector-mediated expression of green fluorescent protein (GFP) was utilized as a novel method for anterogradely tracing the regeneration of brainstem axons. AAV vectors also were used to determine if expression of a developmentally regulated transcription factor, MASH-1, allows mature brainstem neurons to regenerate axons more effectively. Compared to control animals, the expression of MASH-1 in brainstem neurons enhanced the regeneration of noradrenergic axons in a SC bridge and improved hindlimb joint movements. Surprisingly, in contrast to previous studies control animals also exhibited some brainstem axon regeneration. Analyses of the host spinal cord/SC bridge interfaces of control animals revealed variation from sharp to irregular boundaries. At the irregular boundaries of the rostral interface in those animals, the regeneration of axons from noradrenergic neurons and other brainstem populations associated with the presence of long astrocyte processes entering the bridge. Furthermore, the total number of astrocyte processes that entered from the rostral and caudal interfaces associated with improvements in hindlimb joint movements. Previous studies of the SC bridge model implanted pre-gelled mixtures of SCs and Matrigel, in contrast to initially fluid mixtures implanted in this dissertation. Therefore, a direct comparison of these two bridge preparations was performed and demonstrated that initially fluid bridges exhibit increases in both brainstem axon regeneration and the number of astrocyte processes in the bridge. In summary, this work is the first to demonstrate that overexpression of MASH-1 can enhance CNS axon regeneration and that astrocyte processes are important for axon regeneration across a SC bridge. The combination of these intrinsic and extrinsic factors offers a new therapeutic strategy for promoting functional recovery after CNS injury. Advisors/Committee Members: Vance P. Lemmon, Walter G. Bradley, Daniel J. Liebl, Mary Lou King, Mary Bartlett Bunge, Paul J. Reier.

Subjects/Keywords: Transcription Factor; Astrocyte; Spinal Cord Injury; Axon Regeneration; Schwann Cell; Adeno-associated Viral Vector; MASH-1

…Generation of the adeno-associated viral vectors was in part carried out by the Viral Vector Core… …74 CHAPTER 2: THE USE OF AN ADENO-ASSOCIATED VIRAL VECTOR TO ASSESS THE REGENERATION OF… …82 Generation of adeno-associated viral (AAV) vectors..................... 83… …KLF JNK MAG MASH-1 MLC MLR MMP mRNA NCAM NF NGF Adeno-Associated Virus Activating Protein-1… …associated genes.................................................... 54 vii Second messengers…

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APA (6^th Edition):

Williams, R. R. (2010). Intrinsic and Extrinsic Factors for the Regeneration of Brainstem Axons after Spinal Cord Injury. (Doctoral Dissertation). University of Miami. Retrieved from https://scholarlyrepository.miami.edu/oa_dissertations/937

Chicago Manual of Style (16^th Edition):

Williams, Ryan R. “Intrinsic and Extrinsic Factors for the Regeneration of Brainstem Axons after Spinal Cord Injury.” 2010. Doctoral Dissertation, University of Miami. Accessed January 19, 2021. https://scholarlyrepository.miami.edu/oa_dissertations/937.

MLA Handbook (7^th Edition):

Williams, Ryan R. “Intrinsic and Extrinsic Factors for the Regeneration of Brainstem Axons after Spinal Cord Injury.” 2010. Web. 19 Jan 2021.

Vancouver:

Williams RR. Intrinsic and Extrinsic Factors for the Regeneration of Brainstem Axons after Spinal Cord Injury. [Internet] [Doctoral dissertation]. University of Miami; 2010. [cited 2021 Jan 19]. Available from: https://scholarlyrepository.miami.edu/oa_dissertations/937.

Council of Science Editors:

Williams RR. Intrinsic and Extrinsic Factors for the Regeneration of Brainstem Axons after Spinal Cord Injury. [Doctoral Dissertation]. University of Miami; 2010. Available from: https://scholarlyrepository.miami.edu/oa_dissertations/937

14. Dupaty, Léa. Evaluation in vivo de protéines immunorégulatrices dérivées de CTLA-4 et de PD-L1 pour leur capacité à inhiber les réponses immunitaires dans le contexte de la thérapie génique musculaire par AAV : In-vivo evaluation of immunoregulatory proteins derived from CTLA-4 and PD-L1 for their capacity to inhibit immuno responses in a context of AAV muscle gene therapy.

Degree: Docteur es, Sciences de la vie et de la sante, 2018, Normandie

URL: http://www.theses.fr/2018NORMR133

►

La thérapie génique consiste à introduire du matériel génétique dans des cellules dans l’objectif de traiter une pathologie. Le plus souvent, la thérapie génique s’effectue… (more)

▼

La thérapie génique consiste à introduire du matériel génétique dans des cellules dans l’objectif de traiter une pathologie. Le plus souvent, la thérapie génique s’effectue au moyen d’un vecteur viral, transportant le gène jusque dans les cellules cibles. Dans le cas des maladies monogéniques, l’adeno-associated virus (AAV) s’est imposé progressivement comme un vecteur de choix. Son absence de pathogénicité, son large tropisme et sa capacité à transduire des cellules quiescentes sont autant d’avantages comparés à d’autres vecteurs utilisés en thérapie génique. L’utilisation d’AAV est approuvé en Europe pour le traitement d’un déficit rare en lipoprotéine lipase et vient récemment d’être approuvé par les autorités américaines pour le traitement d’un déficit de la vision. Toutefois, les essais de thérapies géniques se heurtent souvent aux réponses immunitaires dirigées contre l’AAV. En effet, les différents composants de ce vecteur viral ont été identifiés comme pouvant déclencher des réponses immunitaires s’opposant à l’efficacité à long terme de la thérapie génique. De plus, la protéine transgénique peut s’avérer immunogène, ce qui conduit au déclenchement de réponses immunitaires, à la destruction des cellules transduites et in fine à l’échec de la thérapie génique. En clinique, des immunosuppresseurs sont utilisés pour palier à ses effets indésirables. Toutefois, de par leurs effets secondaires infectieux et tumorigènes, des stratégies visant plutôt à induire de la tolérance vis-à-vis de la protéine transgénique, associées à un bénéfice pour la santé des patients, se sont développées.L’objectif de ce travail de thèse a été d’implémenter une nouvelle stratégie visant à étudier l’effet immunorégulateur et tolérogène de protéines de fusion dérivées de CTLA-4/Fc et de PD-L1/Fc. Pour cela, nous avons utilisé un modèle murin récapitulant les réponses immunitaires induites par un AAV permettant l’expression d’une protéine modèle fortement immunogène, l’ovalbumine (Ova). Ensuite, des AAV codant pour les protéines au potentiel immunorégulateur ont été synthétisés et injectés aux souris conjointement à l’AAV-Ova. Cette stratégie d’immunorégulation vectorisée (VIR) nous a permis d’évaluer la capacité de chacune des protéines à moduler les réponses immunitaires dirigées contre l’Ova directement in vivo. Au total, ce travail a permis de mettre en évidence i) l’intérêt et les limites de la stratégie VIR, ii) celui du rôle délétère de CTLA-4/Fc au long terme sur les lymphocytes Tregs CD4+FoxP3+, périphériques et centraux, iii) et de démontrer l’intérêt de 2 nouvelles molécules dérivées PD-L1/Fc sur la persistance de l’Ova.

Gene therapy consist into introducing genetic material into cells to treat genetic disorders. Most gene therapies use viral vectors to carry the gene within target cells. In case of monogenic disorders, adeno-associated viruses (AAV) has become a vector of choice because of its lack of pathogenicity, its large tropism and its capacity to transduce quiescent cells. The use of AAV is approved in Europe to…

Advisors/Committee Members: Adriouch, Sahil (thesis director).

Subjects/Keywords: Thérapie génique; Point de contrôle immunologique; Tolérance immunitaire; CTLA-4; PD-L1; Vecteur adéno-associé; Vectorisation; Gene Therapy; Immune checkpoint; Immune tolerance; CTLA-4; PD-L1; Adeno-associated vector; Vectorization; 615.8; 616.079

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Dupaty, L. (2018). Evaluation in vivo de protéines immunorégulatrices dérivées de CTLA-4 et de PD-L1 pour leur capacité à inhiber les réponses immunitaires dans le contexte de la thérapie génique musculaire par AAV : In-vivo evaluation of immunoregulatory proteins derived from CTLA-4 and PD-L1 for their capacity to inhibit immuno responses in a context of AAV muscle gene therapy. (Doctoral Dissertation). Normandie. Retrieved from http://www.theses.fr/2018NORMR133

Chicago Manual of Style (16^th Edition):

Dupaty, Léa. “Evaluation in vivo de protéines immunorégulatrices dérivées de CTLA-4 et de PD-L1 pour leur capacité à inhiber les réponses immunitaires dans le contexte de la thérapie génique musculaire par AAV : In-vivo evaluation of immunoregulatory proteins derived from CTLA-4 and PD-L1 for their capacity to inhibit immuno responses in a context of AAV muscle gene therapy.” 2018. Doctoral Dissertation, Normandie. Accessed January 19, 2021. http://www.theses.fr/2018NORMR133.

MLA Handbook (7^th Edition):

Dupaty, Léa. “Evaluation in vivo de protéines immunorégulatrices dérivées de CTLA-4 et de PD-L1 pour leur capacité à inhiber les réponses immunitaires dans le contexte de la thérapie génique musculaire par AAV : In-vivo evaluation of immunoregulatory proteins derived from CTLA-4 and PD-L1 for their capacity to inhibit immuno responses in a context of AAV muscle gene therapy.” 2018. Web. 19 Jan 2021.

Vancouver:

Dupaty L. Evaluation in vivo de protéines immunorégulatrices dérivées de CTLA-4 et de PD-L1 pour leur capacité à inhiber les réponses immunitaires dans le contexte de la thérapie génique musculaire par AAV : In-vivo evaluation of immunoregulatory proteins derived from CTLA-4 and PD-L1 for their capacity to inhibit immuno responses in a context of AAV muscle gene therapy. [Internet] [Doctoral dissertation]. Normandie; 2018. [cited 2021 Jan 19]. Available from: http://www.theses.fr/2018NORMR133.

Council of Science Editors:

Dupaty L. Evaluation in vivo de protéines immunorégulatrices dérivées de CTLA-4 et de PD-L1 pour leur capacité à inhiber les réponses immunitaires dans le contexte de la thérapie génique musculaire par AAV : In-vivo evaluation of immunoregulatory proteins derived from CTLA-4 and PD-L1 for their capacity to inhibit immuno responses in a context of AAV muscle gene therapy. [Doctoral Dissertation]. Normandie; 2018. Available from: http://www.theses.fr/2018NORMR133

University of South Florida

15. Carty, Nikisha Christine. Recombinant AAV Gene Therapy and Delivery.

Degree: 2009, University of South Florida

URL: https://scholarcommons.usf.edu/etd/1890

► Alzheimer's disease (AD), first characterized in the early 20th century, is a common form of dementia which can occur as a result of genetic mutations… (more)

▼ Alzheimer's disease (AD), first characterized in the early 20th century, is a common form of dementia which can occur as a result of genetic mutations in the genes encoding presenilin 1, presenilin 2, or amyloid precursor protein (APP). These genetic alterations can accelerate the pathological characteristics of AD, including the formation of extracellular neuritic plaques composed of amyloid beta peptides and the formation of intracellular neurofibrillary tangles consisting of hyperphosphorylated tau protein. Ultimately, AD results in gross neuron loss in the brain which is evidenced clinically as a progressive decline in mental capacity. A strong body of scientific evidence has previously demonstrated that the driving factor in the pathogenesis of AD is potentially the accumulation of Aß peptides in the brain. Thus, reduction of Aß deposition is a major therapeutic strategy in the treatment of AD. Recently it has been suggested that Aß accumulation in the brain is modulated, not only by Aß production, but also by its degradation. Several important studies have demonstrated that Aß degradation is modulated by several endogenous zinc metalloproteases shown to have amyloid degrading capabilities. These endogenous proteases include neprilysin (NEP), endothelin converting enzyme (ECE), insulin degrading enzyme (IDE) and matrix metalloprotease 9 (MMP9). In this investigation we study the effects of upregulating expression of several of these proteases through administration of recombinant adeno-associated viral vector (rAAV) containing both endogenous and synthetic genes for ECE and NEP on amyloid deposition in amyloid precursor protein (APP) plus presenilin-1 (PS1) transgenic mice. rAAV administration directly into the brain resulted in increased expression of ECE and NEP and a substantial decrease in amyloid pathology. We were able to significantly increase the area of viral distribution by using novel delivery methods resulting in increased gene expression and distribution. These data support great potential of gene therapy as a method of treatment for neurological diseases. Optimization of gene transfer methods aimed at a particular cell type and brain region in the CNS can be accomplished using AAV serotype specificity and novel delivery techniques leading to successful gene transduction thus providing a promising therapeutic avenue through which to treat AD.

Subjects/Keywords: Beta amyloid; amyloid degrading enzyme; convection enhanced delivery; mannitol; adeno-associated viral vector; transgenic mice; American Studies; Arts and Humanities

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Carty, N. C. (2009). Recombinant AAV Gene Therapy and Delivery. (Thesis). University of South Florida. Retrieved from https://scholarcommons.usf.edu/etd/1890

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Carty, Nikisha Christine. “Recombinant AAV Gene Therapy and Delivery.” 2009. Thesis, University of South Florida. Accessed January 19, 2021. https://scholarcommons.usf.edu/etd/1890.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Carty, Nikisha Christine. “Recombinant AAV Gene Therapy and Delivery.” 2009. Web. 19 Jan 2021.

Vancouver:

Carty NC. Recombinant AAV Gene Therapy and Delivery. [Internet] [Thesis]. University of South Florida; 2009. [cited 2021 Jan 19]. Available from: https://scholarcommons.usf.edu/etd/1890.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Carty NC. Recombinant AAV Gene Therapy and Delivery. [Thesis]. University of South Florida; 2009. Available from: https://scholarcommons.usf.edu/etd/1890

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

The Ohio State University

16. Likhite, Shibi B. Therapeutic suppression of mutant SOD1 by AAV9-mediated gene therapy approach in Amyotrophic Lateral Sclerosis.

Degree: PhD, Molecular, Cellular and Developmental Biology, 2014, The Ohio State University

URL: http://rave.ohiolink.edu/etdc/view?acc_num=osu1417394084

► Amyotrophic Lateral Sclerosis is one of the most common, adult-onset neurodegenerative disorder, characterized by progressive and fatal loss of motor neurons in spinal cord, motor… (more)

Subjects/Keywords: Biology; Biomedical Research; Immunology; Molecular Biology; Neurosciences; Neurobiology; Neurology; Virology; Amyotrophic Lateral Sclerosis, ALS; Superoxide Dismutase 1, SOD1; Adeno-associated vector 9, AAV9; short hairpin RNA, shRNA; Astrocytes; Neuroinflamation; Galectin 1, Gal1; Microglia

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Likhite, S. B. (2014). Therapeutic suppression of mutant SOD1 by AAV9-mediated gene therapy approach in Amyotrophic Lateral Sclerosis. (Doctoral Dissertation). The Ohio State University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=osu1417394084

Chicago Manual of Style (16^th Edition):

Likhite, Shibi B. “Therapeutic suppression of mutant SOD1 by AAV9-mediated gene therapy approach in Amyotrophic Lateral Sclerosis.” 2014. Doctoral Dissertation, The Ohio State University. Accessed January 19, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=osu1417394084.

MLA Handbook (7^th Edition):

Likhite, Shibi B. “Therapeutic suppression of mutant SOD1 by AAV9-mediated gene therapy approach in Amyotrophic Lateral Sclerosis.” 2014. Web. 19 Jan 2021.

Vancouver:

Likhite SB. Therapeutic suppression of mutant SOD1 by AAV9-mediated gene therapy approach in Amyotrophic Lateral Sclerosis. [Internet] [Doctoral dissertation]. The Ohio State University; 2014. [cited 2021 Jan 19]. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1417394084.

Council of Science Editors:

Likhite SB. Therapeutic suppression of mutant SOD1 by AAV9-mediated gene therapy approach in Amyotrophic Lateral Sclerosis. [Doctoral Dissertation]. The Ohio State University; 2014. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1417394084

University of Florida

17. Li, Hong. Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency.

Degree: PhD, Pharmaceutical Sciences - Pharmacy, 2009, University of Florida

URL: https://ufdc.ufl.edu/UFE0024912

► Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency Alpha 1-antitrypsin (AAT) deficiency is a genetic defect caused mostly by a single base substitution in… (more)

▼ Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency Alpha 1-antitrypsin (AAT) deficiency is a genetic defect caused mostly by a single base substitution in the AAT gene, and leads to hepatocyte dysfunction or lung destruction. Protein replacement therapy is the only available treatment for AAT deficiency associated lung disease and requires weekly repeated intravenous infusion of human AAT (hAAT) protein. For AAT deficiency associated liver disease, no effective therapy is available except liver organ transplantation which is limited by the shortage of donor organ. Recent studies showed that adult stem cell gene therapy which replaces the patients? disease-causing gene with the healthy counterparts in their own stem cells holds great potential for the treatment of genetic diseases. To test the feasibility of adult stem cell-mediated liver gene therapy for treatment of AAT deficiency, we performed a series of experiments using three types of adult stem cells including liver progenitor cells (oval cells), bone marrow (BM) cells and adipose tissue-derived mesenchymal stem cells (AT-MSCs) for ex vivo transduction and transplantation. Using oval cells, we confirmed the feasibility of recombinant adeno-associated virus (rAAV) vector mediated ex vivo transduction and transplantation. Considering isolation of oval cell for autologous transplantation is not clinically applicable, we have test the use of BM cells. We showed that both lentiviral vector and rAAV vectors can transduce BM cells. Transplantation of transduced BM cells showed that BM cells transdifferentiated into hepatocytes and mediated transgene (hAAT) expression in the liver. Importantly, sustained serum levels of hAAT were detected in the recipient mice. Similarly, we have employed AT-MSCs since they can be obtained easily (or less invasively) in large quantities. Results from this study demonstrated that AT-MSCs were transduced efficiently by rAAV serotype 1 vector. After transplantation, these cells engrafted into recipient liver, transdifferentiated into hepatocytes, contributed to liver regeneration, and served as platform for transgene expression. Sustained serum levels of hAAT in the recipients implied a potential application for future treatment for AAT deficiency. AT-MSC-based gene therapy presents a novel approach for the treatment of human genetic diseases, such as AAT deficiency. Future studies will focus on achieving therapeutic levels of transgene expression, and gene correction in AT-MSCs for AAT deficiency associated liver disease. ( en ) Advisors/Committee Members: Song, Sihong (committee chair), Hughes, Jeffrey (committee member), Hochhaus, Guenther (committee member), Petersen, Bryon E. (committee member).

Subjects/Keywords: Adipose tissues; Adult stem cells; Bone marrow; Cells; Ellipses; Gene therapy; Hepatocytes; Liver; Stem cells; Transgenes; 1, adeno, adipose, adult, alpha, antitrypsin, associated, bone, cell, deficiency, derived, hepatic, lentiviral, marrow, mesenchymal, oval, stem, tissue, vector, virus

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Li, H. (2009). Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency. (Doctoral Dissertation). University of Florida. Retrieved from https://ufdc.ufl.edu/UFE0024912

Chicago Manual of Style (16^th Edition):

Li, Hong. “Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency.” 2009. Doctoral Dissertation, University of Florida. Accessed January 19, 2021. https://ufdc.ufl.edu/UFE0024912.

MLA Handbook (7^th Edition):

Li, Hong. “Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency.” 2009. Web. 19 Jan 2021.

Vancouver:

Li H. Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency. [Internet] [Doctoral dissertation]. University of Florida; 2009. [cited 2021 Jan 19]. Available from: https://ufdc.ufl.edu/UFE0024912.

Council of Science Editors:

Li H. Adult Stem Cell-Based Gene Therapy For Alpha 1-Antitrypsin Deficiency. [Doctoral Dissertation]. University of Florida; 2009. Available from: https://ufdc.ufl.edu/UFE0024912

Open Access Theses and Dissertations