K. Reed Clark, Ph.D.

K. Reed Clark, PhD is principal investigator in the Center for Gene Therapy at The Research Institute at Nationwide Children’s Hospital and Associate Professor in the Department of Pediatrics at The Ohio State University College of Medicine.

Languages Spoken:

• English

Research Center:

• Center for Gene Therapy

Areas of Interest:

• My laboratory is interested in utilizing recombinant adeno-associated virus mediated (rAAV) gene transfer to develop meaningful treatments for genetic and infectious diseases. Current research focuses on three principle areas: (i) use of antibody gene transfer as a means to augment host immunity; (ii) use of RNA based anti-sense therapeutics to achieve exon-skipping of the dystrophin gene for treatment of DMD; and (iii) development of scalable rAAV production platforms using novel AAV serotypes to support clinical trials. Antibody gene transfer: The focus of this project is to develop AAV delivery vectors that efficiently express antibodies of predetermined specificity within host tissue. Because of the inherent flexibility of this system, light and heavy chain antibody genes can be incorporated into a single rAAV vector, and the antibody-expressing vector can then be used to transduce target tissue in vivo (muscle, liver, and pleural cavity). This in turn, leads to sustained expression of biologically active antibody molecules in the circulation. We have shown that human anti-HIV-1 monoclonal antibodies can be expressed in mice and non-human primates – endowing the host with anti-HIV-1 in vitro neutralization activity. Thus, this approach allows for predetermination of antibody affinity and specificity prior to "immunization", and avoids the need for an active humoral immune response. Beyond the current application for HIV-1 infection, this strategy might be useful for other situations where antibodies with predetermined specificities need to be delivered in vivo. Currently, we are evaluating methods to further augment in vivo levels using various AAV serotypes and routes of delivery. Exon-Skipping: The goal of this project is to develop a meaningful therapy to treat muscular dystrophy caused by mutations within the dystrophin gene that ablate the reading frame. We have constructed several rAAV gene transfer vectors that express anti-sense splice sequences linked to a modified U7 small nuclear RNA (snRNA). Expression of these RNAs block inclusion of the mutated dystrophin exon into the mRNA transcript. This results in the generation of a functional (albeit slightly smaller) dystrophin mRNA and protein product. Currently, we are evaluating exonic splice enhancers (ESEs) as potential targets for efficient skipping and methods of vascular delivery to the muscles of the lower limbs. Lastly, we are developing scalable production and purification methods using stable cell lines and transient transfection technologies for the production of multiple AAV serotypes (1-9). Current research has focused on developing a cGMP “friendly” approach to large-scale production that can be transitioned into our production facility upon its completion.

Undergraduate School

• The Ohio State University
  Date Completed: 06/30/1984

Post Doctoral

• The Ohio State University
  Date Completed: 06/30/1992

2005–present

• Associate Center Director, Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, OH

2002–present

• Associate Professor, Department of Pediatrics, Division of Molecular Medicine, The Ohio State University, Columbus, OH

1997–present

• Director, Viral Vector Core Facility, The Research Institute at Nationwide Children's Hospital, Columbus, OH

1997–present

• Assistant Professor, Department of Pediatrics, Division of Molecular Medicine, The Ohio State University, Columbus, OH

1997–2002

• Assistant Professor , Department of Pediatrics and Molecular Virology and Molecular Genetics, Division of Molecular Medicine, The Ohio State University, Columbus, OH

1992–1996

• Post-Doctoral Fellow, Laboratory of Philip R. Johnson, M.D., Department of Pediatrics, Division of Molecular Medicine, The Ohio State University, Columbus, OH

1988–1990

• Lecturer, Introductory Genetics, The Ohio State University, Columbus, OH

1986–1990

• Graduate Teaching Assistant in Molecular Genetics, The Ohio State University, Columbus, OH

• Rosales XQ, Malik V, Sneh A, Chen L, Lewis S, Kota J, Gastier-Foster JM, Astbury C, Pyatt R, Reshmi S, Rodino-Klapac LR, Clark KR, Mendell JR, Sahenk Z. 2013. Impaired regeneration in LGMD2A supported by increased PAX7-positive satellite cell content and muscle-specific microrna dysregulation.  MUSCLE & NERVE. Vol. 47, no. May: 731-739.
• Heller, KN, Montgomery, CL, Janssen, PML, Clark, KR, Mendell, JR, Rodino-Klapac L. 2013. AAV-mediated overexpression of human a7 integrin leads to histological and functional improvement in dystrophic mice.  Mol Ther. Vol. 21, no. 3. (March): 520-525.
• Wallace, L, Moreo, A, Clark, KR, Harper, SQ. 2013. Dose-dependent Toxicity of Humanized Renilla reniformis GFP (hrGFP) Limits Its Utility as a Reporter Gene in Mouse Muscle.  Mol Ther Nucleic Acids. Vol. 2, no. January: ee86.
• Mendell JR, Rodino-Klapac L, Sahenk Z, Malik V, Kaspar BK, Walker CM, Clark KR. 2012. Gene therapy for muscular dystrophy: Lessons learned and path forward.  Neuroscience Letters. Vol. 527, no. May: 90-9.
• Clark,K,Reed; Penaud-Budloo,Magalie. 2011. Evaluation of the fate of rAAV genomes following in vivo administration.  Methods in molecular biology (Clifton, N.J.). Vol. 807, no. January: 239-258.
• Ganesan LP, Mohanty S, Kim J, Clark KR, Robinson JM, Anderson CL. 2011. Rapid and Efficient Clearance of Blood-borne Virus by Liver Sinusoidal Endothelium.  PLoS Pathogens. Vol. 7, no. 9. (January): ee1002281.
• Clark KR, Penaud-Budloo M. 2011. Evaluation of the Fate of rAAV Genomes Following In Vivio Administration.  Methods Mol Biol. Vol. 807, no. January: 239-258.
• Mendell,Jerry,R; Campbell,Katherine; Rodino-Klapac,Louise; Sahenk,Zarife; Shilling,Chris; Lewis,Sarah; Bowles,Dawn; Gray,Steven; Li,Chengwen; Galloway,Gloria; Malik,Vinod; Coley,Brian; Clark,K,Reed; Li,Juan; Xiao,Xiao; Samulski,Jade; McPhee,Scott,W; Samulski,R,Jude; Walker,Christopher,M. 2010. Brief Report: Dystrophin Immunity in Duchenne's Muscular Dystrophy.  NEW ENGLAND JOURNAL OF MEDICINE. Vol. 363, no. 15. (October): 1429-1437.
• Lock,Martin; McGorray,Susan; Auricchio,Alberto; Ayuso,Eduard; Beecham,E,Jeffrey; Blouin-Tavel,Veronique; Bosch,Fatima; Bose,Mahuya; Byrne,Barry,J; Caton,Tina; Chiorini,John,A; Chtarto,Abdelwahed; Clark,K,Reed; Conlon,Thomas; Darmon,Christophe; Doria,Monica; Douar,Anne; Flotte,Terence,R; Francis,Joyce,D; Francois,Achille; Giacca,Mauro; Korn,Michael,T; Korytov,Irina; Leon,Xavier; Leuchs,Barbara; Lux,Gabriele; Melas,Catherine; Mizukami,Hiroaki; Moullier,Philippe; Mueller,Marcus; Ozawa,Keiya; Philipsberg,Tina; Poulard,Karine; Raupp,Christina; Riviere,Christel; Roosendaal,Sigrid,D; Samulski,R,Jude; Soltys,Steven,M; Surosky,Richard; Tenenbau,Liliane; Thomas,Darby,L; van Montfort,Bart; Veres,Gabor; Wright,J,Fraser; Xu,YiLi; Zelenaia,Olga; Zentilin,Lorena; Snyder,Richard,O. 2010. Characterization of a Recombinant Adeno-Associated Virus Type 2 Reference Standard Material.  HUMAN GENE THERAPY. Vol. 21, no. 10. (October): 1273-1285.
• Mendell JR, Rodino-Klapac, L Rosales XQ, Coley BD, Galloway G, Lewis S, Malik V, Shilling C, Byrne BJ, Conlon T, Campbell KJ, Bremer WG, Taylor LE, Flanigan KM, Gastier-Foster JM, Astbury C, Kota J, Sahenk Z, Walker CM, Clark KR. 2010. Sustained alpha-sarcoglycan gene expression after gene transfer in limb-girdle muscular dystrophy, type 2D.  Annals of Neurology. Vol. 68, no. January: 629-638. PubMed ID: 21031578
• Rodino-Klapac,Louise,R; Montgomery,Chrystal,L; Bremer,William,G; Shontz,Kimberly,M; Malik,Vinod; Davis,Nancy; Sprinkle,Spencer; Campbell,Katherine,J; Sahenk,Zarife; Clark,K,Reed; Walker,Christopher,M; Mendell,Jerry,R; Chicoine,Louis,G. 2010. Persistent Expression of FLAG-tagged Micro-dystrophin in Nonhuman Primates Following Intramuscular and Vascular Delivery.  MOLECULAR THERAPY. Vol. 18, no. 1. (January): 109-117.
• Mendell JR, Campbell K, Rodino-Klapac L, Sahenk Z, Shilling C, Lewis S, Bowles D, Gray S, Li C, Galloway G, Malik V, Coley B, Clark KR, Li J, Xiao X, Samulski J, McPhee SW, Samulski RJ, Walker CM. 2010. Dystrophin Immunity in Duchenne’s Muscular Dystrophy.  New England Journal of Medicine. Vol. 363, no. January: 1429-1437. PubMed ID: 20925545
• Rodino-Klapac LR; Montgomery CL; Bremer WG; Shontz KM; Malik V; Davis N; Sprinkle S; Campbell KJ; Sahenk Z; Clark KR; Walker CM; Mendell JR; Chicoine LG. 2010. Persistent expression of FLAG-tagged micro dystrophin in nonhuman primates following intramuscular and vascular delivery.  Molecular Therapy: The Journal Of The American Society Of Gene Therapy. Vol. 18, no. 1. (January): e109.
• Lock M, McGorray S, Auricchio A, Ayuso E, Beecham EJ, Blouin V, Bosch F, Bose M, Byrne B, Caton T, Chiorini J, Chtarto A, Clark KR, Conlon T, Darmon C, Doria M, Douar AM, Flotte TR, Francis J, Francois A, Giacca M, Korn M, Korytov I, Leon X, Leuchs B, Lux G, Melas C, Mizukami H, Moullier P, Muller M, Ozawa K, Philipsberg T, Poulard K, Raupp C, Riviere C, Roosendaal S, Samulski RJ, Soltys S, Surosky R, Tenenbaum L, Thomas DL, van Montfort B, Veres G, Wright JF, Xu Y, Zelenaia O, Zentilin L, Snyder RO. 2010. Characterization of a Recombinant Adeno-Associated Virus Type 2 Reference Standard Material.  Human Gene Therapy. Vol. 21, no. 10. (January): 1273-1285. PubMed ID: 20486768
• Kota J; Handy CR; Haidet AM; Montgomery CL; Eagle A; Rodino-Klapac LR; Tucker D; Shilling CJ; Therlfall WR; Walker CM; Weisbrode SE; Janssen PM; Clark KR; Sahenk Z; Mendell JR; Kaspar BK. 2009. Follistatin gene delivery enhances muscle growth and strength in nonhuman primates.  Science Translational Medicine. Vol. 1, no. 6. (November): e6ra15.
• Kota,Janaiah; Handy,Chalonda,R; Haidet,Amanda,M; Montgomery,Chrystal,L; Eagle,Amy; Rodino-Klapac,Louise,R; Tucker,Danielle; Shilling,Christopher,J; Therlfall,Walter,R; Walker,Christopher,M; Weisbrode,Steven,E; Janssen,Paul,ML; Clark,K,Reed; Sahenk,Zarife; Mendell,Jerry,R; Kaspar,Brian,K. 2009. Follistatin Gene Delivery Enhances Muscle Growth and Strength in Nonhuman Primates.  SCIENCE TRANSLATIONAL MEDICINE. Vol. 1, no. 6. (November): e6ra15.
• Mendell,Jerry,R; Rodino-Klapac,Louise,R; Rosales-Quintero,Xiomara; Kota,Janaiah; Coley,Brian,D; Galloway,Gloria; Craenen,Josepha,M; Lewis,Sarah; Malik,Vinod; Shilling,Christopher; Byrne,Barry,J; Conlon,Thomas; Campbell,Katherine,J; Bremer,William,G; Viollet,Laurence; Walker,Christopher,M; Sahenk,Zarife; Clark,K,Reed. 2009. Limb-Girdle Muscular Dystrophy Type 2D Gene Therapy Restores alpha-Sarcoglycan and Associated Proteins.  ANNALS OF NEUROLOGY. Vol. 66, no. 3. (September): 290-297.
• Mendell JR; Rodino-Klapac LR; Rosales-Quintero X; Kota J; Coley BD; Galloway G; Craenen JM; Lewis S; Malik V; Shilling C; Byrne BJ; Conlon T; Campbell KJ; Bremer WG; Viollet L; Walker CM; Sahenk Z; Clark KR. 2009. Limb-girdle muscular dystrophy type 2D gene therapy restores alpha-sarcoglycan and associated proteins.  Annals Of Neurology. Vol. 66, no. 3. (September): e290.
• Johnson PR; Schnepp BC; Zhang J; Connell MJ; Greene SM; Yuste E; Desrosiers RC; Clark KR. 2009. Vector-mediated gene transfer engenders long-lived neutralizing activity and protection against SIV infection in monkeys.  Nature Medicine. Vol. 15, no. 8. (August): e901.
• Kota J; Chivukula RR; O'Donnell KA; Wentzel EA; Montgomery CL; Hwang HW; Chang TC; Vivekanandan P; Torbenson M; Clark KR; Mendell JR; Mendell JT. 2009. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model.  Cell. Vol. 137, no. 6. (June): e1005.
• Martin PT; Xu R; Rodino-Klapac LR; Oglesbay E; Camboni M; Montgomery CL; Shontz K; Chicoine LG; Clark KR; Sahenk Z; Mendell JR; Janssen PM. 2009. Overexpression of Galgt2 in skeletal muscle prevents injury resulting from eccentric contractions in both mdx and wild-type mice.  American Journal Of Physiology. Cell Physiology. Vol. 296, no. 3. (March): eC476.
• Schnepp BC; Jensen RL; Clark KR; Johnson PR. 2009. Infectious molecular clones of adeno-associated virus isolated directly from human tissues.  Journal Of Virology. Vol. 83, no. 3. (February): e1456.
• Clark KR, Walsh ST. 2009. Crystal structure of a 3B3 variant-A broadly neutralizing HIV-1 scFv antibody.  Protein Science. Vol. 18, no. 12. (January): 2429-2441. PubMed ID: 19785005
• Rodino-Klapac LR; Lee JS; Mulligan RC; Clark KR; Mendell JR. 2008. Lack of toxicity of alpha-sarcoglycan overexpression supports clinical gene transfer trial in LGMD2D.  Neurology. Vol. 71, no. 4. (July): e240.
• Rodino-Klapac LR; Janssen PM; Montgomery CL; Coley BD; Chicoine LG; Clark KR; Mendell JR. 2007. A translational approach for limb vascular delivery of the micro-dystrophin gene without high volume or high pressure for treatment of Duchenne muscular dystrophy.  Journal Of Translational Medicine. Vol. 5, no. January: e45.
• Mendell JR; Clark KR. 2006. Risks, benefits, and consent in the age of gene therapy.  Neurology. Vol. 66, no. 7. (April): e964.
• Mendell JR and Clark KR. 2006. Challenges for gene therapy for muscular dystrophy.  Curr Neurol Neurosci Rep. Vol. 6, no. 1. (January): 47-56. PubMed ID: 16469271
• Chen CL; Jensen RL; Schnepp BC; Connell MJ; Shell R; Sferra TJ; Bartlett JS; Clark KR; Johnson PR. 2005. Molecular characterization of adeno-associated viruses infecting children.  Journal Of Virology. Vol. 79, no. 23. (December): e14781.
• Schnepp BC; Jensen RL; Chen CL; Johnson PR; Clark KR. 2005. Characterization of adeno-associated virus genomes isolated from human tissues.  Journal Of Virology. Vol. 79, no. 23. (December): e14793.
• Johnson PR; Schnepp BC; Connell MJ; Rohne D; Robinson S; Krivulka GR; Lord CI; Zinn R; Montefiori DC; Letvin NL; Clark KR. 2005. Novel adeno-associated virus vector vaccine restricts replication of simian immunodeficiency virus in macaques.  Journal Of Virology. Vol. 79, no. 2. (January): e955.
• van den Pol AN; Acuna-Goycolea C; Clark KR; Ghosh PK. 2004. Physiological properties of hypothalamic MCH neurons identified with selective expression of reporter gene after recombinant virus infection.  Neuron. Vol. 42, no. 4. (May): e635.
• Chotani MA; Mitra S; Su BY; Flavahan S; Eid AH; Clark KR; Montague CR; Paris H; Handy DE; Flavahan NA. 2004. Regulation of alpha(2)-adrenoceptors in human vascular smooth muscle cells.  American Journal Of Physiology. Heart And Circulatory Physiology. Vol. 286, no. 1. (January): eH59.
• Wang C; Wang CM; Clark KR; Sferra TJ. 2003. Recombinant AAV serotype 1 transduction efficiency and tropism in the murine brain.  Gene Therapy. Vol. 10, no. 17. (August): e1528.
• Schnepp BC; Clark KR; Klemanski DL; Pacak CA; Johnson PR. 2003. Genetic fate of recombinant adeno-associated virus vector genomes in muscle.  Journal Of Virology. Vol. 77, no. 6. (March): e3495.
• Sun Y; Jin K; Clark KR; Peel A; Mao XO; Chang Q; Simon RP; Greenberg DA. 2003. Adeno-associated virus-mediated delivery of BCL-w gene improves outcome after transient focal cerebral ischemia.  Gene Therapy. Vol. 10, no. 2. (January): e115.
• Reber KM; Su BY; Clark KR; Pohlman DL; Miller CE; Nowicki PT. 2002. Developmental expression of eNOS in postnatal swine mesenteric artery.  American Journal Of Physiology. Gastrointestinal And Liver Physiology. Vol. 283, no. 6. (December): eG1328.
• Lewis AD; Chen R; Montefiori DC; Johnson PR; Clark KR. 2002. Generation of neutralizing activity against human immunodeficiency virus type 1 in serum by antibody gene transfer.  Journal Of Virology. Vol. 76, no. 17. (September): e8769.
• Clark, KR. 2002. Recent advances in recombinant adeno-associated virus vector (rAAV) production.  Kidney International. Vol. 61, no. January: S9-S15. PubMed ID: 11841606
• Schnepp B and Clark KR. 2002. Highly purified recombinant adeno-associated virus vectors. Preparation and quantitation.  Methods Mol Med.. Vol. 69, no. January: 427-443. PubMed ID: 11987793
• Clark KR and Johnson PR. 2001. Gene Delivery for Vaccines for Infectious Diseases.  Curr Opin Mol Ther.. Vol. 3, no. 4. (August): 375-384. PubMed ID: 11525561
• Su B; Mitra S; Gregg H; Flavahan S; Chotani MA; Clark KR; Goldschmidt-Clermont PJ; Flavahan NA. 2001. Redox regulation of vascular smooth muscle cell differentiation.  Circulation Research. Vol. 89, no. July: 39-46.
• Liu X; Voulgaropoulou F; Chen R; Johnson PR; Clark KR. 2000. Selective Rep-Cap gene amplification as a mechanism for high-titer recombinant AAV production from stable cell lines.  Molecular Therapy: The Journal Of The American Society Of Gene Therapy. Vol. 2, no. 4. (October): e394.
• Sferra TJ; Qu G; McNeely D; Rennard R; Clark KR; Lo WD; Johnson PR. 2000. Recombinant adeno-associated virus-mediated correction of lysosomal storage within the central nervous system of the adult mucopolysaccharidosis type VII mouse.  Human Gene Therapy. Vol. 11, no. 4. (March): e507.
• Clark KR; Sferra TJ; Lo W; Qu G; Chen R; Johnson PR. 1999. Gene transfer into the CNS using recombinant adeno-associated virus: analysis of vector DNA forms resulting in sustained expression.  Journal Of Drug Targeting. Vol. 7, no. 4. (December): e269.
• Carson WE, Yu H, Dierksheide J, Pfeffer K, Bouchard P, Clark R, Durbin J, Baldwin AS, Peschon J, Johnson PR, Ku G, Baumann H, and Caligiuri MA. 1999. A Fatal Cytokine-Induced Systemic Inflammatory Response Reveals a Critical Role for NK Cells.  Journal of Immunology. Vol. 8, no. 162. (April): 4943-4951.
• Clark KR; Liu X; McGrath JP; Johnson PR. 1999. Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses.  Human Gene Therapy. Vol. 10, no. 6. (April): e1031.
• Liu XL; Clark KR; Johnson PR. 1999. Production of recombinant adeno-associated virus vectors using a packaging cell line and a hybrid recombinant adenovirus.  Gene Therapy. Vol. 6, no. 2. (February): e293.
• Lo WD, Qu G, Sferra TJ, Clark R, Chen R, Johnson PR. 1999. Adeno-associated virus mediated gene transfer to the brain: duration and modulation of expression.  Human Gene THerapy. Vol. 2, no. 10. (January): 201-213.
• Nahman NS Jr; Clark KR; Sferra TJ; Urban KE; Troike AE; Kronenberger J; Sedmak DD. 1998. Successful DNA transfer in cultured human mesangial cells using replication deficient recombinant adenovirus.  Journal Of Investigative Medicine: The Official Publication Of The American Federation For Clinical Research. Vol. 46, no. 5. (June): e204.
• Clark KR; Sferra TJ; Johnson PR. 1997. Recombinant adeno-associated viral vectors mediate long-term transgene expression in muscle.  Human Gene Therapy. Vol. 8, no. 6. (April): e659.
• Clark KR; Voulgaropoulou F; Johnson PR. 1996. A stable cell line carrying adenovirus-inducible rep and cap genes allows for infectivity titration of adeno-associated virus vectors.  Gene Therapy. Vol. 3, no. 12. (December): e1124.
• Clark KR; Voulgaropoulou F; Fraley DM; Johnson PR. 1995. Cell lines for the production of recombinant adeno-associated virus.  Human Gene Therapy. Vol. 6, no. 10. (October): e1329.
• Clark KR; Okuley JJ; Sims TL. 1995. Complete nucleotide sequence of the S1-RNase gene of Petunia hybrida.  Plant Physiology. Vol. 107, no. 1. (January): e307.
• Clark KR; Sims TL. 1994. The S-ribonuclease gene of Petunia hybrida is expressed in nonstylar tissue, including immature anthers.  Plant Physiology. Vol. 106, no. 1. (September): e25.
• Clark KR; Okuley JJ; Collins PD; Sims TL. 1990. Sequence variability and developmental expression of S-alleles in self-incompatible and pseudo-self-compatible petunia.  The Plant Cell. Vol. 2, no. 8. (August): e815.
• Rosales, XQ, Malik, Sheh, A, Chen, L, Lewis S, Kota, Gastier-Foster, JM, Astbury C, Pyatt R, Reshmi S,Rodino-Klapac, LR, Clark, KR, Mendell, JR, Sahenk, Z. Impaired regeneration in LGMD 2A supported by increased Pax7 positive satellite cell content and muscle specific microRNA dysregulation.  Muscle and Nerve.