Research Scientist

Curriculum-Vitae

Amaresh Kumar Ranjan, Ph.D.

Cardiology, 3-33 Atran Building, Contacts:

Mount Sinai School of Medicine, Mobile: 347-***-**** One Gustave L. Levy Place E-mails: *************@*****.*** New York, NY 10029 *******.******@****.***

USA

Objective:

Amaresh Ranjan’s career objective is to understand the molecular and cellular cues during different disease conditions and apply the gained knowledge to devise successful therapies. Peer Reviewed Publications:

1. Hardikar AA, Satoor SN, Karandikar MS, Joglekar MV, Puranik AS, Wong W, Kumar S, Limaye A, Bhat DS, Januszewski AS, Umrani MR, Ranjan AK, Apte K, Yajnik PC, Bhonde RR, Galande S, Keech AC, Jenkins AJ and Yajnik CS. (2015) Susceptibility to obesity and diabetes following multigenerational undernutrition is not reversed by two generations of nutrient availability. Cell Metabolism (Cover story) – Vol. 22 Iss. 2, August 4. Commentaries/News thus far on this original article - 1. Charles Moore: People From Developing Countries More Susceptible To Obesity, Type 2 Diabetes And Cardiovascular Disease Than Western Caucasians Due To Ancestors' Diets. July 17th, 2015. http://diabetesnewsjournal.com/

2. Sonali Prasad: Wolf of Food Street : India’s Growing Nutritional Crisis. Huffington Post India. July 17, 2015.

3. Radio program by Dr. Norman Swan: The epigenetic factors of obesity. Monday 13 July 2015, 5:50PM, http://www.abc.net.au/.

4. IANS/Sydney: Why Indians are at higher risk of diabetes. The Indian Express. July 11, 2015. 5. Ancestral Diet Determine Vulnerability to Type 2 Diabetes. http://ausfoodnews.com.au/ . July 15, 2015.

2. Shapiro SD*, Ranjan AK*, Kawase Y, Cheng RK, Kara RJ, Bhattacharya R, Guzman-Martinez G, Sanz J, Garcia MJ, Chaudhry HW. (2014) Cyclin A2 Induces Cardiac Regeneration After Myocardial Infarction Through Cytokinesis of Adult Cardiomyocytes. Science Translational Medicine 19;6(224). *Equal contributions.

Commentaries thus far on this original article -

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1. Daniele Torella, Ciro Indolfi, and Bernardo Nadal-Ginard : Generation of new cardiomyocytes after injury: de novoformation from resident progenitors vs. replication of pre-existing cardiomyocytes. Ann Transl Med (Editorials). 2015 May; 3(Suppl 1): S8. 2. Maron DF: Gene Therapy Shows Promise For Treating Heart Attack Victims. Scientific American, February 19, 2014.

3. Lu S: Gene therapy discovery may help the heart heal itself, TEDMED Blog Catalyst Series, February 19, 2014.

4. Mullin E: Gene therapy regenerates damaged tissue in pigs after heart attack, Fierce Biotech Research, February 20, 2014.

5. MRC Clinical Sciences Centre Biomedical Picture of the Day, March 18, 2014- Healing Hearts: http://bpod.mrc.ac.uk/archive/2014/3/18

6. Fidler B: With Gene Therapy, VentriNova Aims to Mend Broken Hearts, Xconomy, March 4, 2014: http://www.xconomy.com/new-york/2014/03/04/with-gene-therapy-ventrinova-aims-to-mend- broken-hearts/

7. Fidler B: East Coast Biotech Roundup: VentriNova, RainDance, Biogen, & More, Xconomy, March 7, 2014: http://www.xconomy.com/boston/2014/03/07/east-coast-biotech-roundup-ventrinova- raindance-biogen-more/

8. Haas M: Heart cells: no longer undivided, Science-Business Exchange (SciBX), March 10, 2014. 3. Ranjan AK, Joglekar MV and Hardikar AA (2014) Characterizing Cellular Identity at One Cell Resolution in In Situ Hybridization Methods, Neuromethods book series, vol. 99 eds Giselbert Hauptmann, Springer Press, NY, USA

4. Taylor CJ, Satoor SN, Ranjan AK, Pereira e Cotta MV, Joglekar MV. (2012) A protocol for measurement of noncoding RNA in human serum. Exp Diabetes Res. 2012:168368 5. Ranjan AK, Joglekar MV, Atre AN, Patole M, Bhonde RR, Hardikar AA. (2012) Simultaneous imaging of microRNA or mRNA territories with protein territory in mammalian cells at single cell resolution. RNA Biology. 9(7):949-53

6. Ranjan AK, Joglekar MV, Atre AN, Patole M, Bhonde RR, Hardikar AA. (2012) Cellular detection of multiple antigens at single cell resolution using antibodies generated from the same species. J Immunol Methods. 379(1-2):42-7.

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7. Kumar U, Ranjan AK, Sharan C, Hardikar AA, Poddar P, Pundle A. (2012) Green Approach Towards Size Controlled Synthesis of Biocompatible Antibacterial Metal Nanoparticles in Aqueous Phase Using Lysozyme. Curr. Nanosc. 2012, 8, 130-140.

8. Ranjan AK, Ashutosh A Hardikar, Anandwardhan A Hardikar (2010) A New Method of Coating Vascular Prostheses with Endothelial Cells. Heart Lung and Circulation. 19(8):496-496. 9. Ranjan AK, Kumar U, Hardikar AA, Poddar P, Nair PD, Hardikar AA. (2009) Human blood vessel-derived endothelial progenitors for endothelialization of small diameter vascular prosthesis. PLoS One 4(11):e7718.

10. Ranjan AK, Joglekar MV and Hardikar AA. (2009) Endothelial cells in pancreatic islet development and function Islets 1(1); 2 – 9.

11. Nagarajan M, Kumar N, Nishanth G, Haribaskar R, Paranthaman K, Gupta J, Mishra M, Vaidhegi R, Kumar S, Ranjan AK, Kumar S (2009) Microsatellite markers of water buffalo, Bubalus bubalis--development, characterisation and linkage disequilibrium studies BMC Genet.10:68. 12. Joglekar MV, Ranjan AK and Hardikar AA (2009) Pancreas development and regeneration. Stem cells: Organogenesis and Cancer Research Signpost/ Transworld Research Network press

(http://www.ressign.com/). Frontiers in Bioscience, NIH, USA. Techniques developed by Amaresh Ranjan:

1. A new method to isolate adult porcine cardiomyocytes and their in vitro culture. (Published in Science Translational Medicine 19;6(224).

2. Combined technique of In situ PCR and immunofluorescence for simultaneous transcript and protein analysis in single cells (Published in RNA Biology 9(7):949-53). 3. New technique for antibody stripping from animal cells to detect multiple antigens at single cell resolution using antibodies generated from the same species. (Published in J Immunol Methods. 379(1-2):42-7). 4. Preparation of a new chemical mixture named “Acidified Glycerol” and development of a new methodology for changing hydrophobicity of artificial blood prostheses made up of ePTFE (expanded Tetra Fluoro Ethylene). (Published in PLoS One 4(11):e7718 and special featured in Plos One Surface Treatments, Page 15 http://www.plosone.org/browse/Surface+treatments?). 5. A new method for coating artificial blood vessels with human endothelial cells. (Published in Heart Lung and Circulation. 19(8):496-496).

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Manuscript in communication:

• Hakami N, Ranjan AK, Hardikar AA, Dusting GJ and Peshavariya H. Protective Role of NADPH oxidase 4 in Human Endothelial Progenitor Cells.

Manuscript under preparation:

• Ranjan AK*, Bolli P*, Joska T*, Saunders J, Vardagrasso C, Bernstein E, Belden W, Chaudhry HW: The Epigenetic Modifiers Underlying Heritable Silencing of Cardiomyocyte CCNA2, in preparation for submission to Science *equal contributors

• Ranjan AK and Chaudhry HW. Adult Cardiac Regeneration by Fibroblast Reprogramming. (Review). Patent:

• Hardikar AA*, Ranjan AK*, Joglekar MV, Jenkins AJ and Keech AC. A signature for endothelial damage in diabetes. (Patent filing is under process, University of Sydney, Australia). * Inventors. Oral / Poster presentation:

• Ranjan AK, Vadakke-Madathil S, Ram A, and Chaudhry HW: Cyclin A2 Mediates Reprogramming In Adult Mammalian Cardiomyocytes. Poster Presentation at NYSTEM Annual Scientific Meeting 2015.

• Vadakke-Madathil S, Ranjan AK, and Chaudhry HW: Novel Role of Placental Cdx-2 Cells in Cardiomyogenesis. Poster Presentation at NYSTEM Annual Scientific Meeting 2015.

• Shapiro SD,*Ranjan AK,* Kawase Y, Cheng RK, Kara RJ, Bhattacharya R, Guzman-Martinez G, Sanz J, Garcia MJ, Chaudhry HW : Healing The Broken Heart With Cyclin A2. Poster presentation at 13th Annual Wound Healing: Science & Industry Symposium (2014) San Juan, Puerto Rico.

• Ranjan AK, Chaudhry HW : Cyclin A2 Induces Cell-cycle Re-entry In Adult Mammalian Cardiomyocytes. Oral and poster presentations at 1st Philadelphia Regional Biomedical Postdoctoral Research Symposium (2013) University of Pennsylvania, Philadelphia.

• Shapiro S*, Ranjan AK*, Kawase Y, Kara RJ, Bhattacharya R, Martinez GG, Sanz J, Garcia MJ, Chaudhry HW: CCNA2 Gene Transfer Induces Cardiac Regeneration And Restores Cardiac Function In A Porcine Model Of Myocardial Infarction. Poster Presentation at NYSTEM 2013: Collaboration & Renewal meeting. * Equal contribution

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• Ranjan AK and Hardikar AA, Generation of Vascular Grafts using Human Endothelial Progenitor Cells, Poster presentation at the 6th International Society for Stem Cell Research (ISSCR), June 2008,Philadelphia, USA.

Conferences / Symposia:

•Ninth Annual Translational Stem Cell Research Conference (2014), New York Stem Cell Foundation

(NYSCF), New York, USA. •The NHLBI Symposium on Cardiovascular Regenerative Medicine (2013), National Institute of Health (NIH) Bethesda, USA. •Eighth Annual Translational Stem Cell Research Conference (2013), New York Stem Cell Foundation (NYSCF), New York, USA. •Seventh Annual Translational Stem Cell Research Conference (2012), New York Stem Cell Foundation (NYSCF), New York, USA. •New York City (NYC) Emerging Technologies Summit (May, 2012), NYU Langone Medical Center. New York, USA.

•Sixth Annual Translational Stem Cell Research Conference (2011), New York Stem Cell Foundation

(NYSCF), New York, USA.

•12th ADNAT Convention, Symposium on “The Biology of Embryonic and Adult Stem Cells” (2008), Centre for Cellular and Molecular Biology (CCMB), Hyderabad, India. Workshops and trainings:

•Training course in “Recombinant DNA Techniques for Postgraduates and Faculty of Medical Biochemistry” (2002), School of Biotechnology, Madurai Kamaraj University. •Training course in

“Radioimmunoassay for Thyroid hormones and ELISA Test for Hepatitis B & HIV” (2000), Tata Main Hospital, Jharkhand. •Training course in “vaccine for Anthrax & Rani Khet disease production through Chick Embryo Culture” (1999), Institute of Animal Health & Production, Patna, Government of Bihar, India.

•Training course in “Detection of Leishmania donovani infection in Kala-azar patients’ blood samples by Polymerase Chain Reaction (PCR)” (1999), organized by Sen Diagnostic Laboratory Pvt. Ltd, Patna, Bihar, India. •Workshop on “Industrial Fermentation Technology” (1998), Modern Food Industries, Ranchi, Jharkhand, India.

Awards and Honours:

1. Editorial Board Member “EC Nutrition Journal” (2016) 2. Travel award to attend 13th Annual Wound Healing: Science & Industry Symposium (2014) San Juan, Puerto Rico.

3. “Disease Model & Mechanism (DMM) Travel Fellowship” from “The Company of Biologists Ltd”, United Kingdom (2010) to visit and work at O’Brien Institute, Melbourne, VIC, Australia for 2 months. 4. Advanced Research Fellowship for completion of on going PhD program from the Council of Scientific and Industrial Research (CSIR), Government of India (2008-2010) 6

5. “International Society of Stem Cell Research (ISSCR) Travel Award” for poster presentation in 6th Annual Meeting of ISSCR at Pennsylvania Convention Center Philadelphia, PA USA (June 11-14, 2008). 6. Prestigious award of Junior Research Fellowship from Council of Scientific and Industrial Research (CSIR), Government of India, to initiate Life science research work in India for a period of 3years (2005-2008). 7. Nominated for well renowned Shyama Prasad Mukherjee (SPM) national level award from Council of Scientific and Industrial Research (CSIR), Government of India (2004). 8. Awarded for National Eligibility for Lectureship from Council of Scientific and Industrial Research (CSIR), Government of India (2003).

9. Department of Biotechnology (DBT) Studentship from Department of Biotechnology (DBT), Government of India, was awarded for pursuing Master of Science (MSc.) in Biotechnology for a period of two years

(2001-2003).

10. Awarded “Special Merit Certificate” of National Service Scheme (NSS), Ministry of Youth affairs and Sports, Government of India, for performing community services and participating in special camping program aimed for Literacy and Plantation in Indian village area (1999-2000). Technical expertise (in brief):

Cell Culture and Differentiation: (a) Adult human cardiomyocyte isolation and culture. (b) Isolation of neonatal and adult mice cardiomyocytes by using Langendorff perfusion system (c) Adult, neonatal and embryonic murine cardiomyocyte culture. (d) Adult porcine cardiomyocyte isolation and culture. (e) Isolation and culture of mouse placental cells. (f) Human Embryonic Stem cell culture and differentiation into cardiomyocytes. (g) Human vascular Endothelial Progenitor Cells (EPCs) isolation from arteries and veins and in vitro expansion, differentiation and cryopreservation. (h) Rolling culture of endothelial cells (i) Human Fetal Pancreatic Islets Progenitors culture and their differentiation into islet hormone producing cells. (j) Culture of tumorogenic and non-tumorogenic cell lines from human ovaries. (k) Insect cell line culture (Sf21) (l) Plant tissue culture (tobacco and mung bean). (m) Human (Hela) and mouse (C2C12) cell lines culture maintenance and cryopreservation. (n) 3D culture of adult mouse cardiomyocytes. Cell Physiology: Single cell calcium flux measurements and imaging in isolated and cultured mammalian cardiomyocytes.

Molecular Biology Techniques: (a) Transcript analyses – In situ PCR for single cell transcript analysis, RNA isolation, Quantitative Real time PCR, TaqMan based Low Density Arrays (TLDAs), Construction of cDNA library. (b) Recombinant DNA – Genomic and plasmid DNA isolation; cloning and sub-cloning, Construction of genomic library and Southern blotting. (c) Proteomics – Protein isolation and purification, HPLC, Western blotting, Gel electrophoresis, 2D gel electrophoresis and Mass spectrometry (MS-MS). Microbial Genetics Techniques: - Bacterial culture methodologies including plating, streaking and culturing techniques, Preparation of Competent cells, Transformation by chemical and electroporation methods, Replica plating and Mutagenesis (MNNG as well as UV induced). 7

Immunotechnology: - Antibody generation against specific proteins in Rabbit and Mouse, Enzyme linked immunosorbent assay (ELISA), Immunofluorescence/ immunohistochemistry. Analytical Biochemistry: - Flow Cytometry (FACS), Water Contact Angle measurement, Fourier Transformed Infra-Red (FTIR) spectroscopy.

Microscopy: - Phase contrast, Confocal (Zeiss LSM 510 and LSM 510 Meta), Atomic force microscopy (AFM) and Time Lapse microscopy.

Plant Biotechnology: -Agrobacterium mediated transformation in tobacco plant; Multiple shoots induction in Vigna mungo.

Teaching Experience:

Biochemistry course to undergraduate students at St. Columba’s College, Hazaribagh, India (Visiting lecturer 2001 – 2003)

Mentoring Experience:

1. Archana Ram (Research rotations in years 2012 and 2013). Project – Cyclin A2 Mediates Dedifferentiation and Subsequent Mitosis in Postnatal Cardiomyocytes. Archana entered into the semifinalists list of “Intel Science Talent Search 2014” with this project. At present she is a student at Massachusetts Institute of Technology.

2. Yujin E. Kim (Research rotation in year 2013). Project- Fetal-placental cells for cardiac repair. At present she is a Ph.D. student at Mayo Clinic, Molecular Pathology and Experimental Therapy Biomedical Science program.

3. Girija Kulkarni (Research rotation 2008-2009). Project – Human endothelial cell isolation and culture. Student at University of Melbourne, Australia.

Papers reviewed:

1. Novel insights into affected pathways in an Epidermolysis Bullosa Simplex Dowling-Meara model keratinocyte cell line - implications in BMP- and Wnt-signaling'. BMC Cell Biology, 4 May 2015, Manuscript ID: CEBI-D-15-00016.

2. Activation of GPR40 attenuates chronic inflammation induced impact on pancreatic beta-cells health and function. BMC Cell Biology, 9 May 2013, Manuscript ID: 4498333529892649. 3. Decreased pre-surgical CD34+/CD144+ cell number in patients undergoing coronary artery bypass grafting compared to coronary artery disease-free valvular patients. Journal of Cardiothoracic Surgery, 14 September 2011, Manuscript ID: 3981071336057898.

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Education and Research Experiences:

1997 - 2001 (B.Sc. Biotechnology): Department of Biotechnology, Vinoba Bhave University, Hazaribagh, India.

2001 – 2003 (M.Sc. Biotechnology): School of Biotechnology, Madurai Kamraj University, Madurai, India. (PI- K. Veluthambi, PhD.)

Project: Construction of a co-integrate vector in Agrobacterium tumefaciens to produce marker free transgenic plants.

2003 - 2004 (Project Assistant): Jawaharlal Nehru Center for Advanced Scientific Research (JNCASR), Bangalore, India. (PI- Tapas K. Kundu, PhD.)

Project: Cloning and expression of HDAC4 gene in baculo-virus expression system. 2004 - 2005 (Research Assistant): Center for Cellular and Molecular Biology (CCMB), Hyderabad, India. (PI- Satish Kumar, Ph.D.)

Project: Development of radiation hybrid mapping panel and DNA markers for buffalo genome. 2005 – 2006 (Lab Rotation): National Centre for Cell Science, Pune, India. (PI- Sharmila Bapat, PhD.) Project: Proteomics study of tumorigenic and non-tumorigenic human ovarian cancer cell lines. 2006 - 2011 (Ph.D. in Biotechnology): National Centre for Cell Science, Pune, India. (PI- Anandwardhan A. Hardikar, PhD.)

Project: Evaluation of vascular endothelial cells as an in vitro model for tissue engineering applications. 2012 – Present (Post doctoral fellow): Cardiology, Mount Sinai School of Medicine, New York, USA. (PI- Dr. Hina W Chaudhry)

Project: - Adult mammalian cardiac regeneration through induction of cell cycle re-entry via ectopic Cyclin A2 expression.

Current Lab (Chaudhry Laboratory) Responsibilities: Amaresh Ranjan is involved in all the experimental research works related to Cyclin A2 and cardiac regeneration which includes adult mouse, pig and human cardiomyocyte culture, adenoviral transfection, microscopy, molecular biology techniques to understand the mechanism of cardiac regeneration. Apart from that, he is also working on another project titled “The Epigenetic Modifiers Underlying Heritable Silencing of Cardiomyocyte CCNA2”. He has contributed significantly to this project and a manuscript is currently being submitted to a high impact international journal with Dr. Ranjan as the lead author. During his Ph.D. he also got experience of managing a biotechnology laboratory. Therefore, apart from his involvement in main lab projects, he also helps in managing Chaudhry laboratory with writing papers, designing projects for grants, guidance to graduate or other visiting students, maintenance of lab equipments, and arrangements for all the lab meetings. He also attends important international conferences and gives presentations at such conferences.

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References:

1) Dr. Hina W. Chaudhry

Associate Professor of Medicine

Director, Cardiovascular Regenerative Medicine

Cardiology, Atran 3-34

Mount Sinai Hospital

One Gustave L. levy Place

New York, NY, 10029, The USA

E-mail. – ****.********@****.***

Phone - +1-212-***-****

2) Anandwardhan A. Hardikar, PhD.

Associate Professor and Australian Future Fellow (ARC) Diabetes and Islet biology Group NHMRC Clinical Trials Centre Faculty of Medicine The University of Sydney

Level 6, Medical Foundation Building, 92-94 Parramatta Road, Camperdown, NSW 2050, Australia

+61 2 9562 5071 +61 2 9565 1863

*****.********@***.****.***.**

W: http://www.ctc.usyd.edu.au Lab website: http://www.isletbiology.com/ 3) Milind S. Patole, PhD.

Molecular Biology Laboratory,

National Centre for Cell Science,

Pune University, Ganeshkhind,

Pune, 411007, India.

E-mail: ******@****.***.**

4) Dr.R.R.Bhonde, PhD.

Technical Director

Stempeutics

Manipal Hospital

Bangalore, India

E-mail: ********@*****.***

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