Medical Internal Medicine
Resume:
Chirag Mandavia, M.D., M.S.
Most recent position:
Post-doctoral Fellow
Department of Internal Medicine
University of Missouri School of Medicine
Phone: (91) - 756-***-****
Email: *********@*****.***
EDUCATION
. Kharkov Medical University M.D. 2000 Medicine
Kharkov, Ukraine
. Bowling Green State University M.S. 2003 Molecular
biology
Ohio 43403, USA
. Educational Commission Certificate 2005 Completion
For Foreign Medical
Graduates ECFMG
o Awarded Certificate of Completion and Verification of Medical
Credentials; Certificate number 0-652-088-6; Issued September 23, 2005
[Cleared United States Medical Licensing Exam Series - USMLE Step
1, Step 2 Clinical Knowledge, Step 2 Clinical Skills and also Step
3]
o Medical academic credentials are evaluated by the Educational
Commission for Foreign Medical Graduates (ECFMG) and are equivalent to
degrees offered in accredited institutions in US.
o Member of American Association of Medical Colleges (AAMC) by exam and
professional qualification. AAMC ID 12164229.
AWARDS AND HONORS
. 1981-1990 Certificates of Merit
St. Xavier's, Ahmedabad, India
Awarded multiple certificates of merit for distinction,
conduct and potential
. 1994-1997 Outstanding medical student recognition
Kharkov Medical University, Ukraine
Offered opportunity for fast-track completion of medical
studies
. 2001-2002 Research scholarship
Bowling Green State University, USA
ACADEMIC APPOINTMENTS and WORK EXPERIENCE
. 2001-2003 Graduate Research and Teaching Assistantship
Dept. of Biology, Bowling Green State University, OH
Mutagenesis and protein interaction studies of angiotensin
receptors.
. 2004-2006 Research Assistant/Associate
Gene-Center, Dept. of Biological Sciences, Hunter College
of the City University of New York, NY
RNA-interference and role of protein phosphatases in growth
and development
. 2006-2009 Senior Technician
Dept. of Ophthalmology, Columbia University Medical Center,
NY
Nuclear receptor studies and high-throughput screening to
identify ligands for orphan nuclear receptors
. 2009-2011 Senior Technician
Dept. of Microbiology and Immunology, Columbia University
Medical Center, NY
Genetic manipulation and characterization of the tight-
adherence (tad) locus of dental pathogenic bacteria
POST-DOCTORAL TRAINING
. 2011-2012 Post-doctoral Fellow
Department of Internal Medicine, University of Missouri
School of Medicine, MO
Molecular and translational studies in insulin signaling,
diabetes and cardiovascular disease
MEMBERSHIPS IN PROFESSIONAL ORGRANIZATIONS
. Member of New York Academy of Sciences (NYAS).
. Member of American Society of Microbiology (ASM).
PUBLICATIONS
A. Publications in review and previous publications:
1. Over-nutrition and metabolic cardiomyopathy (Review).
Chirag H. Mandavia, Lakshmi Pulakat, Vincent DeMarco and James R.
Sowers
Metabolism 2011 Oct - PMID: 22565089.
2. Cardiac dysfunction in diabetes (Editorial).
Chirag H. Mandavia, Annayya R. Aroor, Vincent G. DeMarco and James
Sowers
Life Sciences 2012 Mar - Manuscript submitted.
3. Aldosterone induces degradation of insulin receptor substrate 1
protein (IRS-1) in a time and dose-dependent manner in HL-1
cardiomyocytes
Chirag H. Mandavia, Lakshmi Pulakat, Shannon Arnold, Adam Whaley-
Connell and James R. Sowers
Journal of Cardiorenal Medicine - Manuscript in preparation.
4. Putative role of phospho-protein phosphatase PP2A in regulation of the
important insulin-signaling pathway protein insulin receptor substrate
1.
Chirag H. Mandavia and James R. Sowers
Journal of Cardiorenal Medicine 2012 - Manuscript submitted.
5. Mitochondria and oxidative stress in the cardiorenal metabolic
syndrome
Annayya R. Aroor, Chirag Mandavia, Jun Ren, James R. Sowers and
Lakshmi Pulakat
Cardiorenal Med. 2012 May; 2(2):87-109. Epub 2012 Feb 7.
PMID:22619657
6. Letter to the Editor: The need for clarification of the "cardiorenal
metabolic syndrome".
Chirag H. Mandavia and Adam Whaley-Connell
Journal of Cardiorenal Medicine - Manuscript in preparation.
7. Insulin resistance and heart failure: molecular mechanisms (Review).
Annayya R. Aroor, MD, PhD, Chirag H. Mandavia, MD and James R. Sowers,
MD
Heart Failure Clinics 2012 Mar - Manuscript submitted.
8. New concepts in biological role and pharmacological modulation of the
renin-angiotensin-aldosterone axis.
Mandavia CH, Ramdas M, Gul R, Pulakat L.
Molecular and Cellular Endocrinology (2012) - Manuscript in
preparation.
9. AT1R resistance to blockade in the opossum proximal tubule cell due to
variations in the binding pocket
Ravi Nistala, Lakshmi Pulakat, Bradley T. Andersen, Catherine Sinak,
Chirag Mandavia, Thomas Thekkumkara, Robert Speth, Adam Whaley-Connell
and James R Sowers
American Journal of Physiology-Renal Physiology 2012 Jan - In press
(DOI: Paper # F00127/2012).
10. Role of Phe308 in the seventh trans-membrane domain of the AT2
receptor in ligand binding and signaling.
Pulakat L, Mandavia CH, Gavini N
Biochem Biophys Res Commun. 2004 Jul 9; 319(4):1138-43.
PMID: 15194486.
11. Ligand-dependent complex formation between the Angiotensin II receptor
subtype AT2 and Na /H exchanger NHE6 in mammalian cells.
Pulakat L, Cooper S, Knowle D, Mandavia C, Bruhl S, Hetrick M, Gavini
N
Peptides 2005 May; 26(5):863-73. Epub 2005 Jan 21.
PMID: 15808917.
12. Cloning vector pVEX2172, complete sequence
Gen-Bank: JF311902.1
AUTHORS Mandavia, C.H. and Figurski, D.H.
TITLE Direct Submission
JOURNAL Submitted (09-FEB-2011) Microbiology and Immunology,
Columbia University, 701 West 168th Street HHSC 1514A, New
York, NY 10032, USA
13. Plasmid pAA56, complete sequence
Gen-Bank: JF934877
AUTHORS Mandavia, C.H. and Figurski, D.H.
TITLE Direct Submission
JOURNAL Submitted (09-MAY-2011) Microbiology and Immunology,
Columbia University, 701 West 168th Street HHSC 1514A, New
York, NY 10032, US
B. Abstracts presented at conferences:
1. Abstract (2012) 38th Meeting of the International Aldosterone
Conference
Abstract Title: Aldosterone induces degradation of insulin receptor
substrate 1 protein (IRS-1) in a time and dose-dependent manner in HL-
1 cardiomyocytes
Chirag H. Mandavia1, 3, 4, Shannon Arnold1, 3, Adam Whaley-Connell1,
3, 4, James R. Sowers1, 2, 3, 4 and Lakshmi Pulakat1, 3, 4, 5
University of Missouri-Columbia School of Medicine, Departments of
Internal Medicine1, Medical Pharmacology and Physiology2 and Nutrition
and Exercise Physiology5, Diabetes and Cardiovascular Research
Center3, and Harry S Truman Veterans Affair Medical Center4, Columbia,
MO
2. Abstract (2012) Endocrine Society 2012 Meeting in Houston, TX
Abstract Title: A role for GPR-30 in aldosterone-dependent reductions
in IRS-1 in cardiomyocytes; Chirag H. Mandavia, Annayya R. Aroor, Adam
Whaley-Connell, Lakshmi Pulakat, and James R. Sowers; Department of
Internal Medicine, University of Missouri-Columbia School of Medicine.
3. Abstract (2005) Annual Dictyostelium Meeting in Autrans, France -
Chirag Mandavia1, Srividhya Venkatesan1, A Garcia1, Sara Anderson1,
Nicola James1, Robert Salzler1, Jinha Jung1, Hideshi Otsuka2, Julian
Gross2 and Robert P. Dottin1 "Analysis of Phospho-protein Phosphatase
PP2A in growth and development of Dictyostelium discoideum". 1
Department of Biological Sciences, Hunter College of the City
University of New York USA, and 2 Department of Biochemistry, Oxford
University, Oxford, UK.
4. Abstract (2012) presented twice at Life Sciences Week, University of
Missouri-Columbia and Annual Research Day, Harry S Truman Veterans
Memorial Hospital -
Hyder SK, Mandavia C, Sowers JR, Nistala R. Angiotensin-II activation
of Na+/H+ exchanger-3 and mTOR-S6K signaling in the proximal tubule is
improved by Dipeptidyl Peptidase-4 inhibitor; Department of Internal
Medicine, University of Missouri-Columbia School of Medicine,
Columbia, MO.
5. Poster presentation (2008) Association for Research in Vision and
Ophthalmology (ARVO) Florida, US - Abstract Title: Comparison of Co-
repressor Peptides in the Development of a High Throughput Screening
Assay for Small Molecule Agonists of the Retinal Nuclear Receptor,
NR2E3. S. Zimov, C. Mandavia, K. Petrukhin. Ophthalmology, Columbia
University, New York, NY.
6. Abstract (2009) Association for Research in Vision and Ophthalmology
(ARVO) Florida, US - Title: High Throughput Identification of Direct
Target Genes for Photoreceptor-specific Orphan Nuclear Receptor NR2E3.
K. Petrukhin1, C. Mandavia1, F. Fonseca-Galea1, I.P. Chernov2.
1Ophthalmology-Harkness Eye Institute, Columbia, University, New York,
NY; 2Russian Academy of Sciences, Institute of Bioorganic Chemistry,
Moscow, Russian Federation.
7. Abstract (2003) OBASM (Ohio Branch of American Society of
Microbiology).
RESEARCH EXPERIENCE
1. Research work:
a. Research projects undertaken (2001-2003)
Mutagenesis studies to study structure-function relationship of
Angiotensin receptors
My research project on mutagenesis and ligand-binding studies of
the Angiotensin receptor subtype AT2 receptor helped to elucidate
the structure-function relationship of the receptor, and generated
exciting results characterizing the importance of the AT2 receptor
in angiotensin receptor mediated actions and functions.
Specifically, a Phe-Ala mutation at position 308 of the Angiotensin
II AT2 receptor subtype was generated using site-directed
mutagenesis. The DNA was subsequently digested with restriction-
enzymes and in-vitro transcribed to generate mRNA. This was then
micro-injected into Xenopus oocytes and subsequently radio-labeled
ligand probes were used to test the binding affinity of the mutated
AT2 receptor, as compared to the wild-type (WT) AT2 receptor. We
found that the ability of the mutant AT2 receptor to bind ligand
was altered as compared to the wild-type. Paper published. (Paper
published - see "Publications". This work was also presented at
annual meeting of Ohio Branch of American Society of Microbiology
(OBASM) and abstract accepted. (April 2003).
Functional characterization of interactions between angiotensin
receptors and other proteins
This research project was undertaken in breast cancer cells to
highlight the importance of angiotensin not only in cellular growth
and proliferation but also in metabolic signaling pathways, and my
work showed angiotensin receptor is capable of modulating function
through direct protein interactions, in addition to signal
transduction. It also showed the existence of diverse cellular
effects of Angiotensin receptor subtype AT2 in modulating function
in human cells. The human breast cancer cell-line MCF was co-
transfected with AT2 and NHE6 using lipofectamine. The cells were
then lysed and subjected to SDS-PAGE gel electrophoresis to
separate the proteins. These were then transferred to a
nitrocellulose membrane, and co-immunoprecipitation experiments and
Western blot using antibodies raised against either NHE6 or AT2
were performed. We found that AT2 interacts with NHE6 in the
presence of the ligand Angiotensin. (Paper published - see
"Publications Similar experiments were done to check for AT2-
Erb interactions. We found that AT2-Erb interactions also take
place, although in the absence of ligand (Unpublished manuscript).
We also undertook studies involving cell culture of mouse B16
carcinoma cell lines (in vitro) and generation of tumors in mice
through injection of cell-line with subsequent treatment (in vivo
studies) for checking effectiveness of drug. ELISA assays were
aimed at studying elevations of certain markers in animals carrying
tumors. In addition, I worked on yeast 2-hybrid system to study
protein-protein interactions through genetic pathway.
b. Research projects undertaken (2004-2006)
RNA-interference and role of phosphatases in growth and development
In order to characterize the essential role of protein phosphatases
in growth and metabolism in micro-organisms, we employed a novel
method of RNA-interference technology (RNAi) by using short-hairpin
RNA (shRNA) to transiently knock-down genes in order to
characterize their function. My research focused on elucidating the
role of PP2A in the growth and development of the organism
Dictyostelium discoideum. For this purpose, transient knock-down of
PP2A using shRNA constructs (RNAi), as well as PP2A-deficient cells
using knock-in (insertional recombination) and knock-out constructs
(homologous recombination) were created. For knock-down of PP2A,
RNA Interference using shRNA constructs was used on PP2A (serine-
threonine phosphatase) of Dictyostelium Discoideum : Cloning of
RNAi constructs to knock down WT protein phosphatase PP2A of
Dictyostelium discoideum and introduction via double
electroporation of mutant PP2A carrying temperature-sensitive
mutation (site-specific mutagenesis) and RNAi construct into D.
discoideum. We analyzed their effect by looking at phenotypic
expression (growth - development at different temperatures),
protein levels and activity using Western Blots and phosphatase
assays. Radioactivity-based and fluorescence-based phosphatase
assays were used to check the activity of the mutated enzyme as
compared to the WT enzyme. Knock-out cells using homologous
recombination, as well as disrupting the PP2A using knock-in
(insertional recombination) constructs were created, and then
analyzed for phenotype and biochemical activity of the WT PP2A-
deficient cells, to better elucidate the role of PP2A in growth and
development of Dictyostelium, as well as to characterize the
temperature-sensitive mutated PP2A.
c. Research projects undertaken (2006-2009)
Nuclear receptor studies and high-throughput screening to identify
ligands and therefore potential therapeutic targets for the orphan
photoreceptor-specific nuclear receptor Nr2e3
My work in this Translational Research Lab focused on identifying
ligands, and in consequence potential drug targets, for the orphan
nuclear receptor RNR (NR2E3, photoreceptor-specific nuclear
receptor). RNR is considered to be essential for the function of
photoreceptor rods and cones, which are essential for normal
vision, and have been proved to degenerate in many forms of macular
degeneration. My research involved work with Baculovirus-mediated
cloning technology for expression of the protein in insect cells,
checking for the presence of full protein at the DNA and protein
levels using viral DNA PCR and Western Blots respectively; as also
active protein using radio-labeled DNA probes. In addition, I used
sophisticated and complicated techniques for cloning of RNR full-
length protein as well as the RNR-LBD (ligand-binding domain) into
Yeast 2-hybrid system, for subsequent cDNA library screening, to
identify interactions at the genetic level. We purified recombinant
protein using Fast Protein Liquid Chromatography (AKTA FPLC system)
by immuno-affinity, and analyzing protein expression using Western
blots. We employed electrophoretic mobility shift assays to
determine functionality of protein by evaluation of its ability to
bind to known response elements on the genes it regulates, as also
for gene expression regulation studies by studying the ability of
the protein to bind to other response elements on other genes and
regulate their expression. In addition, I was involved in protein
induction and expression studies and several other projects related
to cloning the Bestrophin gene (also known as VMD2) responsible for
Best (or vitelliform) macular dystrophy into the Baculoviral system
for similar studies.
d. Research projects undertaken (2009-2011)
Genetic manipulation and characterization of the tight adherence
(tad) locus of the periodontal pathogen Aggregatibacter
actinomycetemcomitans
Based on my research work in this lab, we were able to identify a
novel method of genetic manipulation in the bacteria
Aggregatibacter actinomycetemcomitans, an important periodontal
pathogen and source of dental caries. In addition, it can also
cause secondary super-infections. Previous research in the lab had
identified and characterized the tight-adherence (tad) locus in
this microorganism, which is crucial for its pathogenicity through
biofilm formation. My research work led to the possibility of using
existing recombineering technology successfully in Aggregatibacter
to facilitate genetic manipulation and the generation of two novel
plasmids, which I have published. These plasmids can be used in a
novel method of recombineering by other researchers intending to do
bacterial manipulations in this organism. In addition, I was
involved in multiple projects involving various studies of,
including, but not limited to, its genetics, and phenotypic
characteristics, and elucidation of some of the 14 genes present in
the tad locus of this organism. In addition to employing
traditional methods, I generated and optimized a modified protocol
for creating gene knockouts and insertions using the recombineering
approach. This work will not only help to characterize the
functional aspects of biofilm formation in Aggregatibacter
actinomycetemcomitans, but will help other researchers interested
in genetic manipulation of bacteria and/or other unrelated genes in
their research and will facilitate an easier and faster methodology
of bacterial genetic manipulation.
e. Research projects undertaken (2011-2012)
Molecular, clinical and translational studies in insulin signaling,
diabetes and cardiovascular disease
This is a clinical and translational Diabetes and Cardiovascular
Research Lab which focuses on diabetes research and insulin
signaling in the heart and other organs and tissues using rodent
models of over-nutrition, insulin resistance and renin-angiotensin-
aldosterone (RAAS) activation. Our lab employs advanced and
sophisticated techniques, such as micro-PET imaging, pressure-
volume loop functional studies, magnetic-resonance imaging (MRI) to
measure cardiac function and glucose uptake and tolerance
measurements by other methods including radioactive glucose uptake
and clamp techniques, among others. Also, focuses on cell signaling
and protein expression and interactions and the role of
mitochondria and oxidative stress involved in insulin signaling and
insulin-mediated glucose uptake using a variety of basic molecular
and immunology techniques such as real-time PCR, immuno-blotting,
ELISA and fluorescence-based assays, cell culture studies,
histological staining, among others.
In addition to being involved in several aspects of the above
lab research, my research here also involves over-expression
studies of the critical insulin signaling pathway molecule insulin
receptor substrate-1 (IRS-1) using green-fluorescent protein (GFP)
tag to analyze and measure its functional effect in the insulin
signaling pathway.
My work on generation of a stable cell line with over-expression
of GFP-tagged IRS-1 in cardiomyocytes will enable to study all
aspects of IRS involvement including:
o Monitoring IRS-1 subcellular localization by fluorescence
microscopy and its migration from the low-density microsomal
fraction (LDM) into the cytosol where it undergoes
degradation
o Involvement of the specific serine residues involved or
preceding in its degradation, and thereby elucidation of the
signaling pathways leading up to this effect, as different
kinases have been shown to serine-phosphorylate different
sites of IRS-1.
o Treatment of cells over-expressing IRS-1 with different
hormones and other effectors involved in the insulin
signaling pathway (for example, components of RAAS such as
angiotensin and aldosterone) will elucidate the mechanistics
of the insulin signaling pathway and IRS-1 signaling in more
detail.
o Protein interaction studies using co-immunoprecipitation
experiments will show binding partners involved in
influencing IRS-1 action.
o In-vitro kinase and phosphatase assays using purified
recombinant IRS-1 as substrate would elucidate the
involvement of the various kinases and phosphatases involved.
2. Techniques and Instrumentation
Knowledge of various techniques used in biotechnology, cellular and
molecular biology, immunology and cancer genetics, including:
1. Fast Protein Liquid Chromatography for protein purification - can use
AKTA FPLC system.
2. Electrophoretic mobility shift assays (EMSAs) to determine ability of
transcription factor proteins to bind to radioactively labeled
response elements on DNA.
3. Baculovirus system cloning, viral DNA isolation and PCR, protein
expression and studies using transfections and Western Blots, cell-
culturing.
4. Metabolic labeling of cells using radioactive ortho-phosphate for in
vivo phosphorylation studies.
5. Laboratory Management - including, but not limited to, ordering,
inventory, lab safety procedures, hazardous/chemical/radioactive
management, invoices and accounting, general routing functions of
laboratory
6. Polymerase Chain Reaction (PCR) for amplification and cloning.
7. Poly-acrylamide gel electrophoresis (SDS-PAGE) with subsequent Western
Blots and co-immunoprecipitation studies, followed by antibody probes
raised against the appropriate proteins.
8. DNA (column and midi-preps, mini-screens etc) and total RNA isolations
with subsequent agarose gel electrophoresis for detection.
9. Transformations of yeast and bacterial E.coli, also transfections into
mammalian cell-lines following cloning into appropriate vectors (tet-
on and tet-off retroviral system and others) or plasmids.
10. Yeast 2-hybrid system: Yeast library screening techniques following
reverse transcription polymerase chain reaction (RT-PCR) techniques to
generate cDNA.
11. Restriction enzyme digestions of plasmid DNA with subsequent in-vitro
transcription to obtain mRNA for micro-injection into frog Xenopus
oocytes (knowledge of Xenopus system) with subsequent radio-active
labeled probes to perform ligand binding assays with use of gamma-
counter and scintillation-counter.
12. Growing and maintaining cell-lines: Growth of B-16 melanoma cell-line
with subsequent injection into mice to generate tumors, followed by
Enzyme-linked immuno-sorbent assay (ELISA) to assess levels of certain
tumor markers.
13. Worked in Animal Facility for 2 years: Handling animals, generating
tumors, treatment, injections etc.
14. Worked in a clinical setting (dealing with patients routinely in the
hospital) during the last 4 years of my undergraduate education in
Medicine.
15. Can work with Xenopus system, radioactivity, handling microscopes,
general lab maintenance, PCR machines, protein gels, can do cell
culture, micro-array hybridization techniques, gene-clean procedures
etc.
16. Constructing shRNA constructs (RNAi) and electroporating into D.
discoideum to knock down specific genes.
17. Constructing knock-out (homologous recombination) and knock-in
constructs (insertional recombination) by cloning techniques for
creating a mutant target gene-deficient cell-line.
3. Bioinformatics knowledge and Computer Skills:
. Expertise in DNA and protein sequence analysis, alignments, related
software packages, generation of primers and probes; familiar with
genotyping, SNPs, gene polymorphisms, structural analysis.
. Packages: Office, Microsoft Word, Excel, Power-point.
. Languages: C, C++, JAVA, Visual Basic 6.0 (Basic knowledge only)
. Operating Systems: DOS (basic knowledge only), Windows XP, 98, 2000,
ME.
Databases: MS Access, SQL, PL/SQL (Basic knowledge only)
Contact this candidate