scholarly journals High-Throughput In Vitro, Ex Vivo, and In Vivo Screen of Adeno-Associated Virus Vectors Based on Physical and Functional Transduction

2020 ◽  
Vol 31 (9-10) ◽  
pp. 575-589 ◽  
Author(s):  
Adrian Westhaus ◽  
Marti Cabanes-Creus ◽  
Arkadiusz Rybicki ◽  
Grober Baltazar ◽  
Renina Gale Navarro ◽  
...  
Keyword(s):  
High Throughput ◽  
Ex Vivo ◽  
Adeno Associated Virus ◽  
Virus Vectors ◽  
In Vivo Screen

scholarly journals Identification and Validation of Small Molecules That Enhance Recombinant Adeno-associated Virus Transduction following High-Throughput Screens

2016 ◽  
Vol 90 (16) ◽  
pp. 7019-7031 ◽  
Author(s):  
Sarah C. Nicolson ◽  
Chengwen Li ◽  
Matthew L. Hirsch ◽  
Vincent Setola ◽  
R. Jude Samulski
Keyword(s):  
Gene Therapy ◽  
Gene Delivery ◽  
High Throughput ◽  
Adeno Associated Virus ◽  
Virus Gene ◽  
Diploid Cells ◽  
High Throughput Screens ◽  
Group 1

ABSTRACTWhile the recent success of adeno-associated virus (AAV)-mediated gene therapy in clinical trials is promising, challenges still face the widespread applicability of recombinant AAV(rAAV). A major goal is to enhance the transduction efficiency of vectors in order to achieve therapeutic levels of gene expression at a vector dose that is below the immunological response threshold. In an attempt to identify novel compounds that enhance rAAV transduction, we performed two high-throughput screens comprising 2,396 compounds. We identified 13 compounds that were capable of enhancing transduction, of which 12 demonstrated vector-specific effects and 1 could also enhance vector-independent transgene expression. Many of these compounds had similar properties and could be categorized into five groups: epipodophyllotoxins (group 1), inducers of DNA damage (group 2), effectors of epigenetic modification (group 3), anthracyclines (group 4), and proteasome inhibitors (group 5). We optimized dosing for the identified compounds in several immortalized human cell lines as well as normal diploid cells. We found that the group 1 epipodophyllotoxins (teniposide and etoposide) consistently produced the greatest transduction enhancement. We also explored transduction enhancement among single-stranded, self-complementary, and fragment vectors and found that the compounds could impact fragmented rAAV2 transduction to an even greater extent than single-stranded vectors.In vivoanalysis of rAAV2 and all of the clinically relevant compounds revealed that, consistent with ourin vitroresults, teniposide exhibited the greatest level of transduction enhancement. Finally, we explored the capability of teniposide to enhance transduction of fragment vectorsin vivousing an AAV8 capsid that is known to exhibit robust liver tropism. Consistent with ourin vitroresults, teniposide coadministration greatly enhanced fragmented rAAV8 transduction at 48 h and 8 days. This study provides a foundation based on the rAAV small-molecule screen methodology, which is ideally used for more-diverse libraries of compounds that can be tested for potentiating rAAV transduction.IMPORTANCEThis study seeks to enhance the capability of adeno-associated viral vectors for therapeutic gene delivery applicable to the treatment of diverse diseases. To do this, a comprehensive panel of FDA-approved drugs were tested in human cells and in animal models to determine if they increased adeno-associated virus gene delivery. The results demonstrate that particular groups of drugs enhance adeno-associated virus gene delivery by unknown mechanisms. In particular, the enhancement of gene delivery was approximately 50 to 100 times better with than without teniposide, a compound that is also used as chemotherapy for cancer. Collectively, these results highlight the potential for FDA-approved drug enhancement of adeno-associated virus gene therapy, which could result in safe and effective treatments for diverse acquired or genetic diseases.

scholarly journals Ex Vivo/In vivo Gene Editing in Hepatocytes Using “All-in-One” CRISPR-Adeno-Associated Virus Vectors with a Self-Linearizing Repair Template

iScience ◽  
2020 ◽  
Vol 23 (1) ◽  
pp. 100764 ◽  
Author(s):  
Simon Alexander Krooss ◽  
Zhen Dai ◽  
Florian Schmidt ◽  
Alice Rovai ◽  
Julia Fakhiri ◽  
...  
Keyword(s):  
Gene Editing ◽  
Ex Vivo ◽  
Adeno Associated Virus ◽  
Virus Vectors ◽  
Repair Template

scholarly journals Strategies for improving the transduction efficiency of single-stranded adeno-associated virus vectors in vitro and in vivo

Gene Therapy ◽  
2008 ◽  
Vol 15 (18) ◽  
pp. 1287-1293 ◽  
Author(s):  
G R Jayandharan ◽  
L Zhong ◽  
B Li ◽  
B Kachniarz ◽  
A Srivastava
Keyword(s):  
Transduction Efficiency ◽  
Adeno Associated Virus ◽  
Virus Vectors

Successful Gene Transfer Using Adeno-Associated Virus Vectors into the Kidney: Comparison among Adeno-Associated Virus Serotype 1–5 Vectors in vitro and in vivo

2004 ◽  
Vol 96 (4) ◽  
pp. e119-e126 ◽  
Author(s):  
Shin-ichi Takeda ◽  
Masafumi Takahashi ◽  
Hiroaki Mizukami ◽  
Eiji Kobayashi ◽  
Koichi Takeuchi ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Adeno Associated Virus ◽  
Virus Vectors ◽  
Virus Serotype ◽  
Serotype 1

scholarly journals Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo

Gene Therapy ◽  
2003 ◽  
Vol 10 (26) ◽  
pp. 2105-2111 ◽  
Author(s):  
Z Wang ◽  
H-I Ma ◽  
J Li ◽  
L Sun ◽  
J Zhang ◽  
...  
Keyword(s):  
Adeno Associated Virus ◽  
Virus Vectors ◽  
Highly Efficient

scholarly journals Adeno-Associated Virus Type 2-Mediated Transduction of Human Monocyte-Derived Dendritic Cells: Implications for Ex Vivo Immunotherapy

2001 ◽  
Vol 75 (19) ◽  
pp. 9493-9501 ◽  
Author(s):  
Selvarangan Ponnazhagan ◽  
Gandham Mahendra ◽  
David T. Curiel ◽  
Denise R. Shaw
Keyword(s):  
Dendritic Cells ◽  
Immune Responses ◽  
Transgene Expression ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Adeno Associated Virus ◽  
Dc Vaccine

ABSTRACT Dendritic cells (DCs) are pivotal antigen-presenting cells for regulating immune responses. A major focus of contemporary vaccine research is the genetic modification of DCs to express antigens or immunomodulatory molecules, utilizing a variety of viral and nonviral vectors, to induce antigen-specific immune responses that ameliorate disease states as diverse as malignancy, infection, autoimmunity, and allergy. The present study has evaluated adeno-associated virus (AAV) type 2 as a vector for ex vivo gene transfer to human peripheral blood monocyte (MO)-derived DCs. AAV is a nonpathogenic parvovirus that infects a wide variety of human cell lineages in vivo and in vitro, for long-term transgene expression without requirements for cell proliferation. The presented data demonstrate that recombinant AAV (rAAV) can efficiently transduce MOs as well as DCs generated by MO culture with granulocyte-macrophage colony-stimulating factor plus interleukin in vitro. rAAV transgene expression in MO-derived DCs could be enhanced by etoposide, previously reported to enhance AAV gene expression. rAAV transduction of freshly purified MO followed by 7 days of culture with cytokines to generate DCs, and subsequent sorting for coexpression of DC markers CD1a and CD40, showed robust transgene expression as well as evidence of nuclear localization of the rAAV genome in the DC population. Phenotypic analyses using multiple markers and functional assays of one-way allogeneic mixed leukocyte reactions indicated that rAAV-transduced MO-derived DCs were as equivalent to nontransduced DCs. These results support the utility of rAAV vectors for future human DC vaccine studies.

Menisci are Efficiently Transduced by Recombinant Adeno-Associated virus Vectors in Vitro and in Vivo

2004 ◽  
Vol 32 (8) ◽  
pp. 1860-1865 ◽  
Author(s):  
Henning Madry ◽  
Magali Cucchiarini ◽  
Gunter Kaul ◽  
Dieter Kohn ◽  
Ernest F. Terwilliger ◽  
...  
Keyword(s):  
Adeno Associated Virus ◽  
Virus Vectors
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