Our Research: Faculty/Research Laboratories

William R.A. Osborne, PhD

Research Professor, Department of Pediatrics

Email Address: wosborne@u.washington.edu

Departmental Website: http://www.washington.edu/medicine/pediatrics/home/

William R. A. Osborne, PhD

Background:

Bill Osborne received his PhD from King’s College, University of London in 1973.  He did post-doctoral training in Dr. Richard Tashian’s laboratory at the University of Michigan.  He is currently a Research Professor at the Diabetes and Obesity Center of Excellence at the University of Washington.

Focus:

Dr. Osborne’s research is focused on virus-mediated gene therapy to treat diabetes and cyclic neutropenia in rat and dog models respectively.  He is Director of the Virus, Molecular Biology and Cell Core of the Diabetes Endocrinology Research Center at the University of Washington. This Core produces lentivirus and retrovirus vectors for affiliated researchers.

Treatment of diabetes by encapsulated cells

Encapsulation of genetically modified cells permits insulin delivery for Type 1 diabetes without the need for immunosuppression.  The long term goals of this work are (1) to develop cloned, well characterized cell lines that may be transplanted to treat rats with diabetes and (2) to develop methods to permit implantation of allogeneic islets for long term survival and therapy without the need for immunosuppression.  This approach has the benefit that only one human cell line will be generated and characterized for the treatment of all patients.  These studies will provide valuable preclinical information from diabetic rodents that may be applied to treat patients with type 1 diabetes.

We are investigating encapsulated cell therapy for the long-term delivery of glucagon like peptide (GLP-1) to treat rat models of type 1 and type 2 diabetes.  GLP-1 exhibits biological and clinical effects that include stimulation of glucose-dependent insulin secretion, insulin biosynthesis, β cell proliferation and neogenesis, inhibition of β cell apoptosis, inhibition of glucagon secretion, slowing of gastric emptying and induction of a satiety signal.

Rats are treated with encapsulated allogeneic vascular smooth muscle cells (VSMC) genetically modified to secrete GLP-1.  In preliminary studies we have shown that allogeneic VSMC transduced to express GLP-1 and delivered to leptin receptor deficient T2DM rats in Theracyte encapsulation devices induced normoglycemia, weight loss and islet proliferation.  Treatment of BB T1D rats by sustained GLP-1 expression before diabetes onset showed plasma GLP-1 levels elevated 3 to 5 fold over control rats. Hypoglycemia was not detected and this was anticipated from the glucose regulated action of GLP-1.  Diabetes onset in untreated rats occurred at 56.5±0.6 SEM (n=6) days of age and in GLP-1 treated T1D rats was delayed until 76.4±3.3 SEM (n=5) days of age (p<0.001).  After disease onset untreated control rats showed rapid weight loss and elevated blood glucose (>650 mg/dl) and did not survive beyond 11 days.  At 5 days after diabetes onset insulin secreting islets were absent in untreated rats.  In contrast, treated T1D rats maintained weight for up to 143 days of age and showed insulin secreting beta cells.  Sustained GLP-1 expression delivered by encapsulated cells before diabetes onset in BB T1D rats showed improved clinical outcome suggesting the potential for treating patients using long lasting GLP-1 analogues.

Lentivirus-mediated transfer of Gimap5 to treat type 1 diabetic rats

The objective of this research is to investigate lentivirus-mediated gene therapy to treat spontaneously diabetic BB rats, a well characterized model of T1DM. The mutant lymphopenia (lyp) gene in BB rats is a novel GTPase immune-associated (Gimap5) protein.  We propose that loss of Gimap5 function in thymocytes of BB rats leads to lymphopenia and autoimmunity and diabetes and hypothesize that lentivirus-mediated Gimap5 expression in thymocytes will delay or reverse diabetes. We will use lentivirus vectors to transduce lyp/lyp bone marrow stem cells to express wild type Gimap5 cDNA under the control of lck proximal promoter, a thymocyte specific promoter. Specific Aim 1: in vitro studies to document transduction efficiency and tissue specific transgene expression targeted at rat bone marrow and T and B cells. In Specific aim 2 we will study in vivo gene expression by transplanting transduced  bone marrow to irradiated recipient rats. The well characterized diabetic BB rat affords us the opportunity to study gene therapy treatment before onset, at onset and after onset when insulin secreting β cells have been destroyed by the autoimmune process. These three stages of intervention in the disease process are applicable to the treatment of T1DM patients.

Treatment of grey collie dogs with cyclic hematopoiesis by lentivirus delivery of G-CSF

Cyclic neutropenia is a rare disease that occurs both in man and grey collie dogs.  The recurrent severe neutropenia leads to bacterial infections and shortened life expectancy.  Daily administration of recombinant G-CSF eliminates the severe recurrent neutropenia and provides therapy for dogs and patients.  The aim of this research is to employ two different approaches to treat grey collie dogs with cyclic neutropenia: virus-mediated G-CSF delivery; and cell therapy using bioisolator devices.  In the first aim we will administer G-CSF-lentivirus to grey collie, normal dogs and rats to determine the duration of vector-mediated cytokine expression, dose response relationships, the ability to re-administer virus and the tissue distribution of virus after administered to the muscle.  In the second aim we will investigate the use of bioisolator devices to deliver sustained therapeutic levels of G-CSF to dogs and rats.  We will also determine if allogeneic cells can be used to deliver G-CSF to rats and normal dogs before treating grey collie dogs. We have successfully treated four dogs with cyclic neutropenia by administration of G-CSF-lentivirus to muscle.  A successfully treated dog has been returned to its owner in Wisconsin and is healthy and living a normal life.  We believe lentivirus-mediated gene expression may be applied to the sustained delivery of glucose regulated insulin in diabetic animals and patients.

Representative Publications:

Barry SC, Ramesh N, Lejnieks DV, Simonson WT, Kemper L, Lernmark A, Osborne WRA: Glucose-regulated insulin expression in diabetic rats. Human Gene Therapy 12:131-139, 2001.

Yanay O, Flint LY, Brzezinski M, Barry SC, Barton RW, Osborne WRA: Long term erythropoietin gene expression form transduced cells in bioisolator devices. Human Gene Therapy 14: 1587-1593, 2003.

Brzezinski M, Yanay O, Waldron L, Barry SC, Osborne WRA. G-CSF-lentivirus administration in rats provided sustained elevated neutrophil counts and subsequent EPO-lentivirus administration increased hematocrits. J Gene Med 9:571-578, 2007.

Sweet IR, Yanay O, Waldron L, Gilbert M, Fuller JM, Tupling T, Lernmark A, Osborne WRA: Treatment of Diabetic Rats with Encapsulated Islets. Journal of Cellular and Molecular Medicine 12: 2644-2650, 2008

Yanay O,Moralejo D, Kernan K, Brzezinski M, Fuller JM, Barton R, Lernmark A. and Osborne, WR. Prolonged survival and improved glycemia in BB diabetic rats after early sustained exposure to GLP-1. J Gene Med 12: 538–544, 2010, DOI: 10.1002/jgm.1466

Moralejo DH, Yanay O, Kernan K, Bailey A, Lernmark A, Osborne W. Sustained glucagon-like peptide 1 expression from encapsulated transduced cells to treat obese diabetic rats.  J Biosci Bioeng. 111. 383-387, 2011 PMID: 21216666

Yanay O, Dale DC, Osborne WR.  Repeated Lentivirus-mediated G-CSF Administration to Treat Canine Cyclic Neutropenia. Human Gene Therapy.. 2012. [Epub ahead of print] PMID:22845776

  • Osborne WRA, Ramesh N, Lau S, Clowes MM, Dale DC, Clowes AD: Gene therapy for long-term expression of erythropoietin in rats. Proc. Natl. Acad. Sci. USA 92:8055-8058, 1995.

    Wei MQ, Lejnieks DV, Ramesh N, Lau S, Seppen J, Osborne WRA: Sustained gene expression in transplanted skin fibroblasts in rats. Gene Therapy 6:840-844, 1999.

    Seppen J, Barry S, Lam GM, Ramesh N, Osborne WRA: Retroviral preparations derived from PA317 packaging cells contain inhibitors that co-purify with viral particles and are devoid of viral vector RNA. Human Gene Therapy 11:771-775, 2000.

    Barry SC, Seppen J, Ramesh N, Foster, JL, Ochs HD, Garcia JV, Osborne WRA: Lentiviral and Murine retroviral transduction of T-cells for expression of human CD40 ligand. Human Gene Therapy 11:323-332, 2000.

    Barry SC, Ramesh N, Lejnieks DV, Simonson WT, Kemper L, Lernmark A, Osborne WRA: Glucose-regulated insulin expression in diabetic rats. Human Gene Therapy 12:131-139, 2001.

    Seppen J, Barry SC, Harder B, Osborne WRA: Lentivirus administration to rat muscle provides efficient sustained expression of erythropoietin. Blood 98:594-596, 2001.

    Barry SC, Harder B, Brzezinski M, Flint LY, Osborne WRA: Lentivirus vectors encoding both central polypurine tract and posttranscriptional regulatory element provide enhanced transduction and transgene expression. Human Gene Therapy 12:1103-1108, 2001.

    Freitas AC, Bento FM, Ramesh N, Osborne WRA, Han SW: Modified basticidin S resistance gene (bsrm) as a selectable marker for construction of retroviral vectors. J Biotechnology 95:57-62, 2002.

    Katen LJ, Aprikyan AG, Dale DC, Osborne WRA: Cloning and sequencing of the canine neutrophil elastase cDNA. DNA Sequence 13:221-223, 2002.

    Yanay O, Barry SC, Katen LJ, Brzezinski M, Flint LY, Christensen J, Liggitt D, Dale DC, Osborne WRA: Treatment of canine cyclic neutropenia by lentivirus-mediated G-CSF delivery. Blood 102:2046-2052, 2003.

    Yanay O, Flint LY, Brzezinski M, Barry SC, Barton RW, Osborne WRA: Long term erythropoietin gene expression form transduced cells in bioisolator devices. Human Gene Therapy 14: 1587-1593, 2003.

    Barry S, Brzezinski M,Yanay O, Seppen JE, Osborne WRA: Sustained elevation of neutrophils in rats induced by lentivirus-mediated G-CSF delivery. J Gene Med 7: 1510-1516, 2005.

    Yanay O, Brzezinski M, Christensen J, Liggitt D. Dale DC. Osborne, WRA: An adult dog with cyclic neutropenia treated by lentivirus-mediated G-CSF delivery. Human Gene Therapy 17: 464-469, 2006.

    Fitzgerald SM, Goyal RK, Osborne WRA, Roy JD, Wilson JW, Ferrell RE: Identification of functional single nucleotide polymorphism haplotypes in the cytidine deaminase promoter. Human Genetics 119:276-283, 2006.

    Brzezinski M, Yanay O, Waldron L, Barry SC, Osborne WRA. G-CSF-lentivirus administration in rats provided sustained elevated neutrophil counts and subsequent EPO-lentivirus administration increased hematocrits. J Gene Med 9:571-578, 2007.

    Yanay, O. IR Sweet, L Waldron, M Brzezinski, ML Gilbert, JM Fuller, T Tupling, A Lernmark, Osborne, W.R.A. L Waldron, M Brzezinski, JM Fuller, T Tupling, EA Rutledge, A Lernmark. Lentivirus-mediated Gimap5 gene delivery to treat diabetic BB rats. Molecular Therapy 15,Supplement 1, S41 2007.

    Tatake R, M. ON, Kennedy C, Hikel V, Runyan V, Monaco K-A, Yu K, Osborne W, Barton R, Schneiderman R: Glucose-regulated insulin production from genetically engineered human non-beta cells. Life Sciences 81:1346-1354, 2007.

    Sweet IR, Yanay O, Waldron L, Gilbert M, Fuller JM, Tupling T, Lernmark A, Osborne WRA: Treatment of Diabetic Rats with Encapsulated Islets. Journal of Cellular and Molecular Medicine 12: 2644-2650, 2008 PMID: 18373735

    Moralejo, D. H., C. T. Hansen, P. Treuting, M. J. Hessner, J. M. Fuller, B. Van Yserloo, R. Jensen, W. Osborne, A. E. Kwitek, and A. Lernmark. Differential effects of leptin receptor mutation on male and female BBDR.Gimap5-/Gimap5- spontaneously diabetic rats. Physiological Genomics 41: 9-20, 2010.

    Yanay O,Moralejo D, Kernan K, Brzezinski M, Fuller JM, Barton R, Lernmark A. and Osborne, WR. Prolonged survival and improved glycemia in BB diabetic rats after early sustained exposure to GLP-1. J Gene Med 12: 538–544, 2010, DOI: 10.1002/jgm.1466.

    Moralejo DH, Yanay O, Kernan K, Bailey A, Lernmark A, Osborne W. Sustained glucagon-like peptide 1 expression from encapsulated transduced cells to treat obese diabetic rats.  J Biosci Bioeng. 111. 383-387, 2011 PMID: 21216666

    Yanay O, Dale DC, Osborne WR.  Repeated Lentivirus-mediated G-CSF Administration to Treat Canine Cyclic Neutropenia. Human Gene Therapy.. Sept 12.  2012. [Epub ahead of print] PMID:22845776

 

Current Collaborations:

Within the Diabetes and Obesity Center of Excellence and its Affiliated Members
Ofer Yanay, MD
Alan Chait, MD
Ian Sweet, PhD
Michael Schwartz, MD

Outside the Diabetes and Obesity Center of Excellence
Ake Lernmark, PhD, Sweden
Daniel Moralejo, DVM, PhD, University of Washington

Lab Members:

Kelly Kernan, BS
Brian Van Yserloo, BS
Julia DeFriel, BS