Program Project Grant

OMB No 0925-0001 and 0925-0002 (Rev 10/15 Approved Through 10/31/2018) ______BIOGRAPHICAL SKETCH NAME POSITION TITLE Associate Dean for Research and Graduate Marcia F. McInerney______Studies (protected time for research) Distinguished University Professor eRA commons user name Medicinal and Biological Chemistry mmciner College of Pharmacy and Pharmaceutical Sciences University of Toledo EDUCATION/TRAINING INSTITUTION AND LOCATION YEAR(s) FIELD OF DEGREE STUDY University of Michigan Ph.D. 1989 Microbiology & Immunology Yale University School of Med. Postdoc 1989-1991 Immunobiology- Dr. Charles A Janeway Jr, Mentor Joslin Diabetes Center/Harvard School of Visiting Prof 1998-1999 Diabetes/ Medicine of Medicine/ Endocrinology Iacocca Fellow University of Michigan Visiting Prof 2008 Metabolism, of Internal Endocrinology, Medicine and Diabetes

A. Personal Statement I have been involved for over 25 years in research on the autoimmune aspects of type 1 diabetes (T1D) and inflammation in type 2 diabetes (T2D). While in the Janeway Laboratory for my postdoctoral we were the first lab to show that islets in nonobese diabetic (NOD) mice did not express MHC class II molecules, but that expression of MHC class I increased significantly during the development of diabetes. Therefore, we established that beta cells were not presenting antigen to CD4 T cells (last publication listed). I have worked actively on autoimmunity in type 1 diabetes as evidenced by publications on autoantigens (insulin, glutamic acid decarboxylase), autoantibodies (anti-insulin receptor), T cells (CD8 and CD4), T cell clones and hybridomas (anti-insulin, antiGAD), humanized transgenic mice expressing the diabetes susceptibility DQ8 molecule that turned out to be an autoimmune model of human myocarditis, and the role of the insulin receptor in diabetes development which was supported by NIH. A co-authored paper in Diabetes explored the role of IL-13 and IL-13R and was accompanied by a commentary. Innate immunity is another area of interest due to my postdoctoral in the Janeway Laboratory. I examined innate immunity, toll-like receptors, and secondary infection (periodontitis) in T1D and showed that bone marrow derived macrophages from diabetic mice are dysfunctional. This work was supported by an NIH grant. In the last 6 years I have been studying inflammation and cytokine involvement in the development of type 2 diabetes with a USDA/NIFA supported grant entitled “Dietary and Genetic Risk Factors in Obesity and Diabetes”. I have a number of papers associated with the metabolic syndrome and a recently published paper in the journal Obesity concerning the interaction between macrophages and adipocytes in obesity and cytokine profiling. A current NIH grant returns to the work on insulin receptor being involved in T cell movement into the pancreas in type 1 diabetes. Future work in the area would involve development of new transgenic models to deliver T regulatory cells to the pancreas. I have been involved with research/teaching of 21 PhD students including 7 as the major advisor and 12 master’s students including 8 as the major advisor. Furthermore, I have worked with an additional 29 undergraduate students on independent research projects making a total of 63 students that I

have mentored over the years. Since 2007 I have been involved in numerous review sessions for NIH and was also an ad Hoc review for the USDA and internationally for Diabetes UK. I served for 5 years on the NIDDK subcommittee B which reviewed K awards and training grants. I have also reviewed postdoctoral fellowship grants for NIH. My research/teaching experience with graduate and undergraduate students and my review service makes my expertise valuable for service on the Advisory board for this grant.

B. Positions and Honors. (Honors in bold) 1988 -1989 Predoctoral Fellowship, American Diabetes Association, University of Michigan "Immunoregulatory Defects in the Nonobese Diabetic (NOD) Mouse." 1989- 1990 Postdoctoral Fellowship, National Institutes of Health Training Grant, Yale University School of Medicine, Charles A. Janeway Jr , Postdoctoral Mentor 1990- 1991 Postdoctoral Fellowship, Juvenile Diabetes Foundation, Yale University School of Medicine, "Regulatory T Cells in Diabetes." Charles A. Janeway Jr., Postdoctoral Mentor 1991 -1997 Assistant Professor, The University of Toledo (UT), College of Pharmacy, Medicinal and Biological Chemistry Department 1993-96 Career Development Award. American Diabetes Association. "Autoantigen Recognition by CD8 T Lymphocytes in Type 1 Diabetes." 1996-97 CODA Children’s Research Award. Central Ohio Diabetes Association. "Upregulated Insulin Receptor Expression on Peripheral T Cells : A Potential Predictor of Diabetes Onset?" 1998 -1999 Senior Mary K. Iacocca Fellow, Joslin Diabetes Center/ Harvard Medical School. Visiting Associate Professor of Medicine, Harvard University and Visiting Scientist, Joslin Diabetes Center. Dr Myra Lipes, mentor 1997 - 2004 Associate Professor, UT College of Pharmacy, Medicinal and Biological Chemistry Dept. Adjunct Associate Professor, UT, Biological Sciences and Pathology Depts. 2004 Promotion to Professor, University of Toledo 2000 - 2013 Interim Chair followed by Chair, Medicinal and Biological Chemistry Dept., College of Pharmacy and Pharmaceutical Sciences, University of Toledo 2006 The University of Toledo Outstanding Faculty Research Award and the Dean’s Award for the College of Pharmacy for Outstanding Research 2008 Visiting Professor of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes. University of Michigan. Dr. Massimo Pietropaolo – mentor. 2008- Adjunct Professor of Internal Medicine, University of Michigan 2010- Distinguished University Professor, University of Toledo 2014 Associate Dean for Research and Graduate Studies (protected time for research) Service 2005- Medical Advisory Board - Diabetes Action Research & Ed Fdn and Grant Reviewer 2007-2012 Grant Review Committee Member NIDDK Subcommittee B; K and Training Grant 2007-11, 2014 Diabetes UK reviewer 2008-14 all ad hoc NIDCR (RFA), NIAID (Program project), NIDDK (2 RFAs), NCI (SEP), and NIDDK Chairman SBIR, STTR 2015 ad hoc USDA/NIFA, NIH IMM-D SEP, NIH IMM-S SEP, NIH F07 Immunology Fellowships 2016 NIH NIDCR 2 review sessions ad hoc, 2 review sessions NIDDK K12 and R01

C. Selected peer-reviewed publications associated with diabetes (out of 31)

Tulsulkar J, Nada SE, Slotterbeck BD, McInerney MF, Shah ZA. Obesity and hyperglycemia lead to impaired post-ischemic recovery after permanent ischemia in mice. Obesity (Silver Spring). 2015 Dec 23. doi: 10.1002/oby.21388. [Epub ahead of print]

Lester SG, Russo L, Ghanem SS, Khuder SS, DeAngelis AM, Esakov EL, Bowman TA, Heinrich G, Al-Share QY, McInerney MF, Philbrick WM, Najjar SM. Hepatic CEACAM1 Over-Expression Protects Against Diet Induced Fibrosis and Inflammation in White Adipose Tissue. Front Endocrinol (Lausanne). 2015 Aug 3;6:116. doi: 10.3389/fendo.2015.00116. eCollection 2015

Lindsey A. Ebke, Andrea L. Nestor-Kalinoski, Brandon D. Slotterbeck, Ali G. Al Dieri, Sumona Ghosh-Lester, Lucia Russo, Sonia M. Najjar, Hermann von Grafenstein, and Marcia F. McInerney. Tight Association between Macrophages and Adipocytes in Obesity: Implications for Adipocyte Preparation. Obesity (Silver Spring). 2014 May;22(5):1246-55

Angevine KR, Wuescher LM, Andrews K, Alexander LA, McInerney MF, Kieffer TJ, Mensah-Osman EJ. Menin and GIP are inversely regulated by food intake and diet via PI3/AKT signaling in the proximal duodenum. Nutr Diabetes. 2012 Dec 3;2:e55.

Rasche SS, Phillips M, McInerney MF, Sercarz, EE and Quinn A. IL-13Ra Expression on Beta Cell- specific T cells in NOD Mice. Diabetes. 2011 Jun;60(6):1716-25. Associated commentary: Wong, FS. Stimulating IL-13 receptors on T cells: a new pathway for tolerance induction in diabetes? pg 1657.

Ghosh S, Kaw M, Patel PR, Ledford KJ, Bowman TA, McInerney MF, Erickson SK, Bourey RE, Najjar SM. Mice with Null Mutation of CEACAM1 Develop Nonalcholic Steatohepatitis. Hepatic Medicine: Research and Evidence 2010; 2: 69–78

Huang S, Kaw M, Harris MT, Ebraheim N, McInerney MF, Najjar SM, Lecka-Czernik B. Decreased Osteoclastogenesis and High Bone Mass in Mice with Impaired Insulin Clearence Due to Liver- Specific Inactivation of CEACAM1. Bone. 2010 Apr; 46(4): 1138-45. Epub 2010 Jan 4

Morran, MP, LA Alexander, B D. Slotterbeck, M F. McInerney. Dysfunctional Innate Immune Responsiveness to P. gingivalis LPS in Diabetes. Oral Microbiology and Immunology 2009; 24(4): 331-339

Quinn, A., McInerney, M.F. Huffman, D., McInerney, B.E., Mayo, S., Haskins, K., and Sercarz, E. T Cells to a Dominant Epitope of GAD65 Express a Public CDR3 Motif. Internat. Immunol. 2006; 18 (6): 967-979.

Mohammad, M., Morran, M. Slotterbeck, B., Leaman, D.W., Sun, Y. vonGrafenstein, H., Hong, S.C., and M.F. McInerney. Dysregulated Toll-like Receptor Expression and Signaling in Bone Marrow- Derived Macrophages at the onset of Diabetes in the Nonobese Diabetic Mouse. Internat. Immunol. 2006; 18 (7): 1101-1113.

McInerney, M.F., Najjar, S.M., Brickley, D., Abou Rjaily, G.A., Lutzke, M., Haskell, B.D., Flurkey, K., Zhang Y-J, Pietropaolo,S.L., Pietropaolo, M., Byers, J.P. and Leiter, E.H. Anti-Insulin Receptor Autoantibodies are Not Required for Type 2 Diabetes Pathogenesis in NZL/Lt Mice, a New Zealand Obese (NZO)- Derived Mouse Strain. Exper. Diabetes Res. 2004;5:177-185.

Havari, E., Lennon-Dumenil, A.M., Taylor, J.A., Turley, S.J., Klein, L., Neely, D., McInerney, M.F., Wucherpfennig, K.W., and Lipes, M.A. Expression of the B7.1 Costimulatory Molecule on Pancreatic Beta Cells Abrogates the Requirement for CD4 T Cells in the Development of Type 1 Diabetes J. Immunol. 2004; 173:787-796.

*Taylor, J.A., *Havari E, McInerney, M.F. Bronson, R.T., Wucherpfennig, K.W. and Lipes, M.A. A Spontaneous Model for Autoimmune Myocarditis Using the Human MHC Molecule HLA-DQ8. J. Immunol. 2004, 172: 2651-2658. (* equivalent first authors)

Quinn, A., M.F. McInerney and E.E. Sercarz. MHC class I- Restricted Determinants on the GAD65 Molecule Induce Spontaneous CTL Activity. J. Immunol. 2001;167:1748-57.

McInerney, M. F., J. Burkey, L. Guan, J. C. Flynn and C. A. Janeway, Jr. An Islet- Specific CD8+ T Cell Hybridoma Generated From Nonobese Diabetic Mice Recognizes Insulin as an Autoantigen. Diabetes Res. & Clin. Prac., 2000;47(3):151-168.

McInerney, M.F., J.C. Flynn, P.J. Goldblatt, S. Najjar, R.S. Sherwin and C.A. Janeway, Jr. High Density Insulin Receptor Positive T Lymphocytes From NOD Mice Transfer Insulitis and Diabetes. J. Immunol., 1996; 157: 3716-3726.

McInerney MF, Rath S, Janeway CA Jr. Exclusive expression of MHC class II proteins on CD45+ cells in pancreatic islets of NOD mice. Diabetes. 1991 May;40(5):648-51.

D. Research Support Current: NIDDK R15: “Chemotaxis to Islets Based on Cellular Insulin Receptor Expression” PI McInerney 7/1/14-6/30/2017

Ohio Board Of Regents Interdisciplinary Grant “Use of a Bioconjugate Vaccine to Alter Autoimmunity in T1D”. 7/1/2013-5/30/2016. PI McInerney. The study is focused on NOD mice and the development of a bioconjugate for delivery of peptides and altered peptide ligands to modulate disease.

Representative Past Completed Grants within the last three years:

2005-2012 USDA/NIFA, Dietary and Genetic Risk Factors in Obesity and Diabetes PI/PD Pharmacy – McInerney, co-PI Dr Sharrel Pinto, PI Medicine Sonia Najjar University of Toledo. This grant combined a basic science investigation on the role of inflammation and the effect of the loss of CEACAM1 in metabolic syndrome with a community outreach portion consisting of medication therapy management for patients with diabetes and hypertension.

Major Scientific Accomplishments Discovery #1: Dr. McInerney discovered when she was a postdoctoral fellow in Dr. Charles Janeway's Laboratory that islet cells in nonobese diabetic (NOD) mice, an animal model for type 1 Diabetes, do not express MHC class II. Only infiltrating cells that were also CD45 positive, of bone marrow origin, expressed MHC class II. This indicated that the islets themselves were not MHC class II antigen presenting cells. This was the primary paper on this topic. Furthermore, the published paper showed that the islet cells in NOD mice had increased expression of MHC class I as diabetes developed in the animal model. Recently it has been shown in humans that islets in type 1 diabetic patients have increased expression of MHC class I. The mechanism for intensified MHC class I expression has not been clarified to date. However, an increase in MHC class I expression can make the islets a better target for autoreactive CD8+ T cells.

McInerney MF, Rath S, Janeway CA Jr. Exclusive expression of MHC class II proteins on CD45+ cells in pancreatic islets of NOD mice. Diabetes. 1991 May;40(5):648-51.

Discovery #2: Graduate work at the University of Michigan supported by the ADA resulted in one of the primary papers on preventing insulitis and diabetes in NOD mice by injection of Complete Freund's Adjuvant (CFA). Most investigators inject peptide or whole antigen emulsified in CFA in order to examine T cell proliferative responses to the antigen. In the case of NOD mice, who spontaneously become diabetic in association with T cell responses to islet antigens, when pathogenic peptides were injected emulsified in CFA NOD mice mounted an innate immune response that lead to blockade of the autoimmune pathway. A component of CFA is mycobacteria. Bacillus- Calmette-Guerin (BCG), an avirulent strain of mycobacteria, is already used as a vaccine for immunoprophylaxis of tuberculosis. Further work by others has shown that BCG induces tumor necrosis factor (TNF) production by stimulating innate immunity. The TNF kills disease-causing autoimmune cells and restores pancreatic beta-cell function through regeneration. Clinical trials are currently underway using BCG to stimulate human innate immunity to restore beta cell function.

McInerney, M.F., S.B. Pek, and D.W. Thomas. Prevention of Insulitis and Diabetes Onset by Treatment with Freund's Adjuvant in NOD mice. Diabetes 1991; 40:715-725.

Discovery #3: This paper sought to determine if CD8+ T cells specific for GAD65 were present in prediabetic NOD mice and whether they were involved in diabetes development. Putative MHC class I (Kd) binding determinants that were proximal to MHC class II determinants were examined for their ability to elicit cytolytic activity. GAD65 peptides 206-214 and 546-554 stimulated naïve spleens cells to produce IFNg and showed antigen specific cytotoxic T lymphocyte activity and lysed targets pulsed with homologous peptide. Furthermore, GAD 546-554 T cells transferred insulitis into NODscid recipients. This published work shows previous experience with GAD peptides and with ELISPOT assays that will be used in this grant. Furthermore, this published paper is evidence for the ability to maintain mice, carry out transfer experiments, and analyze the pancreatic tissue for infiltrating cells. Pertinent to this submission, Dr. Quinn used altered peptide ligands of GAD65 to suppress proinflammatory cytokines made by GAD specific T cell clones from NOD mice. Work on autoantigens also included insulin as an autoantigen in Type 1 Diabetes.

Quinn, A., M.F. McInerney, E.E. Sercarz. MHC class I-Restricted Determinants on the GAD65 Molecule Induce Spontaneous CTL Activity. JI 2001;167:1748-57.

Discovery#4: While working with Dr Charles Janeway Jr, who first described PAMP (Pathogen associated molecular patterns), and then Toll-like receptors were discovered to recognize PAMP bacterial and viral ligands and linked to innate immunity. I examined innate immunity in the NOD mouse model of Type 1 Diabetes and discovered that macrophages are dysfunctional under hyperglycemic conditions and that the same could occur with periodontal infection explaining why T1D patients are susceptible to

periodontitis.

Morran, MP, LA Alexander, B D. Slotterbeck, M F. McInerney. Dysfunctional Innate Immune Responsiveness to P. gingivalis LPS in Diabetes. Oral Microbiology and Immunology 2009; 24(4): 331-339

Discovery #5: While collaborating with Sonia Najjar on the USDa grant we had several papers on the role of CEACAM1 in obesity and type 2 Diabetes. Furthermore, I explored inflammation in obesity by looking at interactions between adipocytes and macrophages. During this latter work we discovered that macrophages adhere tightly to adipocytes and are still adherent after collagenase digestion and washing. Macrophages appear in the floating layer with the adipocytes indicating a floating layer contaminated with macrophages even at single cell level of digestion.

Heinrich G, Russo L, Castaneda TR, Pfeiffer V, Ghadieh HE, Ghanem SS, Wu J, Faulkner LD, Ergün S, McInerney MF, Hill JW, Najjar SM. Leptin resistance contributes to obesity in mice with null mutation of carcinoembryonic antigen cell adhesion molecule 1. J Biol Chem. 2016 Mar 21. pii: jbc.M116.716431. [Epub ahead of print]

Lee SJ, Heinrich G, Fedorova L, Al-Share QY, Ledford KJ, Fernstrom MA, McInerney MF, Erickson SK, Gatto-Weis C, Najjar SM. Development of nonalcoholic steatohepatitis in insulin-resistant liver- specific S503A carcinoembryonic antigen-related cell adhesion molecule 1 mutant mice. Gastroenterology. 2008 Dec;135(6):2084-95. Epub 2008 Aug 20.