BIOMEDICAL SCIENCES GRADUATE PROGRAM FALL 2012

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1 THE OHIO STATE UNIVERSITY BIOMEDICAL SCIENCES GRADUATE PROGRAM FALL 2012 Reeva Aggarwal PhD Candidate MECHANISMS OF HUMAN CD34+ STEM CELL-MEDIATED REGULATION OF OSTEOPOROSIS IN A PRECLINICAL MODEL November 27 th, 2012 DHLRI, Room # :00 AM

2 VITA July, Born Chandigarh, India BS, Biophysics, Panjab University MS, Biophysics, Panjab University COMMITTEE MEMBERS Hiranmoy Das, PhD, Advisor Periannan Kuppusamy, PhD Sudha Agarwal, PhD Nancy L. Lill, PhD

3 ABSTRACT Osteoporosis is a systemic metabolic bone disease characterized by low mineral density and micro architectural deterioration of skeleton. Osteoporosis or porous bone disease can progress asymptomatic until bones fracture. It is a major cause of morbidity and medical expense worldwide. Lack of physical activity, hormonal disturbances, age, gender, hereditary and nutrition are some the factors implicated to cause this disease. Physiologically, bone is a dynamic organ that constantly undergoes remodeling. Bone consists of dense organic, inorganic and mineral components. At cellular level, balance/coupling between the activities of bone forming cells (osteoblasts) and bone resorbing cells (osteoclasts) maintains bone homeostasis. Increased bone loss due to increased osteoclast and decreased osteoblast activities is considered as an underlying cause of osteoporosis. Several therapies such as hormone replacement therapy in women, ingestion of anti-resorptives (bisphosphonates), implantation of osteoinductive device combined with change in lifestyle offers only temporary augmentation of bone mineral density with side effects. Since, cures for osteoporosis are limited, consequently the potential of stem cell based therapies is currently being considered. Adult stem cells, we used for this study express CD133+/CD34 surface marker, and are multipotent in nature, isolated from human umbilical cord blood, could be differentiated into many lineages depending upon the molecular cues received from their microenvironment. However, their limited numbers from a single unit restricts further experimental use of those cells. Thus, our lab developed a nanofiber-based expansion technology to obtain adequate numbers of CD34+ cells isolated for experimental use and potential therapeutic applications. Herein, we show that nanofiber-expanded CD34+ cells could be differentiated into osteoblastic lineage, in vitro. Systemically delivered CD34+ cells home to the bone marrow and significantly improve bone deposition, bone mineral density and bone micro-architecture in osteoporotic aged nonobese diabetic severe combined immunodeficient (NOD/SCID) mice. The elevated levels of osteocalcin, IL-10, GM-CSF, and decreased levels of MCP-1 in serum parallel the improvements in bone micro-architecture. Furthermore, CD34+ cells improved osteoblast activity and concurrently impaired osteoclast differentiation, maturation, and functionality. Mechanistically, CD34+ cells offer osteoprotection via the upregulation of osteoblastic factors such as Runtrelated transcription factor 2 (RUNX2) and its downstream effectors such as collagen type I alpha1 and osteocalcin. Consistent with our in vivo observations of impaired osteoclastic activities after CD34+ stem cell therapy, CD34+ cells have the ability to regulate nuclear translocation of osteoclastogenic differentiation factor called nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), and thus negatively regulating its downstream transcriptional activity, and osteoclastogenesis. Taken together, these findings demonstrate a novel approach for utilizing nanofiber-expanded CD34+ cells as a therapeutic application for the treatment of osteoporosis in a preclinical model.

4 RECENT ABSTRACTS AND PRESENTATION Human T Cells Limit Breast Tumor Growth Partly by Down Regulating Cell Survival-Related Molecules and Up Regulating Apoptosis-Related Molecules in Tumor Cells, AACR Annual Meeting; Chicago, IL, April 2012 Regulation of osteoblasts and osteoclasts by human umbilical cord blood derived CD34 stem cells in osteoporotic NOD/SCID mice. 9th Annual meeting of International Society for Stem Cell Research (ISSCR); Toronto, ON, Canada, June 2011 Preclinical Application of Hematopoietic Stem Cells for Osteoporosis; Finalist in Basic Biological Sciences Category, 25th Annual Edward Hayes Graduate Forum; OSU, Columbus, OH, February 2011 HSCs improve bone mineral density and bone remodelling in murine osteoporotic model. National Center for Regenerative Medicine (NCRM); Case Western Reserve University, Cleveland, OH. November 2010 Osteoblastic differentiation of hematopoietic stem cells for the treatment of osteoporosis. Annual DHLRI & OSUMC Research Day, Columbus, OH. October, 2009 AWARDS AND HONORS Finalist, March th Edward F. Hayes Graduate Research Forum, OSU, Columbus, OH. Best Poster, November Annual Retreat of National Center for Regenerative Medicine (NCRM); Case Western Reserve University, Cleveland, OH.

5 RECENT PUBLICATIONS Original articles Aggarwal R, Lu J, Kanji S, Matthew J, Das M, Noble GJ, McMichael BK, Agarwal S, Hart RT, Sun Z, Lee BS, Rosol TJ, Jackson R, Mao HQ, Pompili VJ and Das H. Human Umbilical Cord Blood-Derived CD34+ Cells Reverse Osteoporosis in NOD/SCID Mice by Altering Osteoblastic and Osteoclastic Activities. PLoS One, 2012, 7(6): e Aggarwal R, Lu J, Kanji S, Matthew J, Das M, Lustberg MB, Ray A, Pompili VJ, Shapiro CL, Das H. Human Vγ2Vδ2 T Cells Limit Breast Cancer Growth by Modulating Cell Survival- and Apoptosis-Related Molecules in Tumor Cells. Submitted Lu J, Aggarwal R, Kanji S, Matthew J, Das M, Pompili VJ and Das H. Human Ovarian Tumor cells Escape γδ T Cell Recognition Partly by Down Regulating Surface Expression of MICA and Limiting Cell Cycle Related Molecules, PLoS One, 2011, 6(9): e23348 Lu J, Kanji S, Aggarwal R, Das, M, Joseph M, Wu LC, Mao HQ, Pompili VJ, Hadjiconstantinou M, and Das H. Umbilical cord-derived hematopoietic stem cells improve dopaminergic neuron morphology in the MPTP-mice. Frontiers in Bioscience, in press. Das, M, Lu J, Joseph M, Aggarwal R, Kanji S, McMichael B, Lee B, Agarwal S, Kuppusamy P, Pompili VJ, Jain M and Das H. Kruppel-like factor 2 (KLF2) Regulates Monocyte Differentiation and Functions in mbsa and IL-1β-Induced Arthritis. Current Molecular Medicine, 2012, 12(2), pp Review articles Aggarwal R, Lu J, Pompili VJ and Das H. Hematopoietic stem cells: transcriptional regulation, ex vivo expansion and clinical application. Current Molecular Medicine, 2012 Jan; 12(1): (Invited Review, Peer-Reviewed). Aggarwal R, Pompili VJ, Das H. Genetic modification of ex-vivo expanded stem cells for clinical application. Frontiers in Bioscience Jun 1; 15: (Invited Review, Peer- Reviewed). Lu J, Aggarwal R, Pompili VJ, Das H. A novel technology for hematopoietic stem cell expansion using combination of nanofiber and growth factors. Recent Patents on Nanotechnology Jun; 4(2): (Invited Review, Peer-Reviewed).

6 Book chapters Aggarwal R, Pompili VJ and Das H. Hematopoietic stem cells and bone regeneration; In Stem Cells and Cancer Stem Cells, Volume 3, M. A. Hayat Ed. Springer in press. Aggarwal R, Pompili VJ and Das H. Hematopoietic stem cells in atherosclerotic development and resolution. In Chronic Inflammation: Molecular Pathophysiology, Nutritional and Therapeutic Interventions. S. Roy, D. Bagchi and S.P. Raychaudhuri Eds. CRC Press 2012: pp (Chapter 6). ISBN: Lu J, Aggarwal R, Pompili VJ and Das H. Ex vivo expanded hematopoietic stem cells for ischemia, In Stem Cells and Cancer Stem Cells, Volume 2, M. A. Hayat Ed, Springer 2012: pp ISBN: Aggarwal R, Lu J, Pompili VJ and Das H. Development of ischemic diseases and stem cell therapy. In Role of Natural Products and Bioactive Compounds in Disease Prevention. M.M. Essa, A. Manickavasagan and E. Sukumar Ed, Nova Science Publishers 2012: pp (Chapter 4). ISBN:

7 Biomedical Sciences Graduate Program 1170 Graves Hall 333 W. 10 th Avenue Columbus, Ohio

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