CURRICULUM VITAE

Rajamani Rathinam Ph.D.

Home Address

**** ******** ****,

Apt # 204, Troy, MI 48084

Phone number: 504-***-****

Email: aczhu3@r.postjobfree.com; aczhu3@r.postjobfree.com Immgration Status: Permanent Resident (Green card) EDUCATION

1994-1997: B.Sc - Biochemistry, University of Madras, Chennai, Tamilnadu, India. 1998-2000: M.Sc-Biochemistry, University of Madras, Chennai, Tamilnadu, India. 2003 to 2008: Ph.D. - Biochemistry-Physiology, University of Madras, Chennai, India. Thesis: Methanol induced free radical generation with protein oxidative damages in folate deficient rat and the role of DL-α-Lipoic acid. Unpaired electron containing atoms or molecules are called free radicals. These radicals are produced by abnormal xenobiotic mechanism in the body and they are responsible for DNA and protein damage. Free radical toxicity is associated with pathophysiology of aging such as cataracts, atherosclerosis, neoplastic diseases, diabetes, diabetic retinopathy, chronic inflammatory diseases of the gastrointestinal tract, aging of skin, diseases associated with cartilage, Alzheimer's disease, and other neurologic disorders. Free radical-induced injury can explain many clinical conditions. Methanol is an adulterant in developing countries, it causes free radical production in body and its toxicity is associated with many clinical conditions. Folate deficiency is major condition to methanol induced free radical formation because loss folate cycle. So it is important to reduce free radical mediated disease. DL-α-Lipoic acid has been reported to effectively reduce oxidative stress. Methanol toxicity affects mainly central nervous system. We studied the effect of the lipoic acid on methanol toxicity in brain regions (cerebral cortex, cerebellum, midbrain, pons medulla, hippocampus and hypothalamus) in rat. To mimic human model we successfully developed folate deficient rat model by folate deficient diet or methotrexate. The level of methanol toxicity was assessed by free radical scavenging enzymes superoxide dismutase, catalase, reduced, oxidized glutathione and lipid-peroxidation in brain and liver samples. It was found that methanol administration increases free radicals, and the lipoic acid supplementation significantly reduces methanol mediated free radical generation. RESEARCH SKILLS

Animal Studies: Handling rat and mice; Implantation of human breast cancer cells into mice for xenograft study. Perfusion and isolation of lungs, liver, kidney and spleen. Subcutaneous and intraperitoneal implantation using cell lines. Lung fixation using Z-fix solution and staining with India ink to analyze lung metastasis in mice. Bone marrow isolation and isolation of mouse embryos. Preparation of mouse embryonic fibroblasts from embryos. Xenograft study using cell lines by breast pad injection (mouse model).Genotyping of animals using PCR. Molecular Biology: PCR techniques, DNA/RNA isolation, sub-cloning of gene fragments into cloning vectors and into mammalian expression vectors. Immunnocytochemistry using cells, Transformation, isolation of plasmids; transient and stable transfection, agarose gel electrophoresis, gel documentation, and experience in High Performance Liquid Chromatography (HPLC) using Refractive Index detector. Have experience in gene knock out study and drug infusion into mice to check cisplatin toxicity. Made retroviral expression of aurora kinase A into fibroblast cells. After cloning Aurora kinase A into retroviral vector, the viral vectors were concentrated in cell culture. Finally made stable cell lines using vector marker by selection. Protein Analysis: Cell lysate preparation, protein assay, western blot analysis, Pull down analysis (using beads) and immunopreciptation. Cell Biology: Cell culture, maintaining transfected cells and plasmid isolation. Preparation of primary cell lines from breast tumors. Detection of apoptosis of cells using flow cytometry with annexin staining. SOFTWARE: MS word, Graph pad prism and gene tool analysis for sequencing results, and restriction mapping.

PREVIOUS APPOINTMENTS

2003-2006: Research Assistant in the Department of Physiology, University of Madras, India under the project entitled “A study of efficacy of alcohol oxidase loaded erythrocytes in combating methanol toxicity” funded by Indian Council of Medical Research (ICMR), New Delhi, India.

Methanol toxicity is initiated by its byproduct formic acid. In this project we tried to eliminate formate by enzyme formate dehydrogenase loaded erythrocytes with carbicarb. Because, formate is metabolized into co2 by formate dehydrogenase. The enzyme was loaded into erythrocytes by hypotonic dialysis method, then the erythrocytes were introduced into methanol intoxicated rats which were already made to folate deficiency with Methotrexate. Blood samples were tested every one hour after erythrocyte administration for four hours. Then blood samples were analyzed for level of formate using HPLC and fluorimetric method and also blood pH, pO2 and pCO2 were analyzed with blood gas analyzer. Finally we found that formate dehydrogenase loaded erythrocytes eliminate formate.

2006-2008: University Research Fellow (URF) in the Department of Physiology, University of Madras, Chennai,India.

2006-2007: Teaching: handled classes to teach Physiology for the post graduate students in the department of Physiology, University of Madras, India. 2008-2011: Postdoctoral Fellow in the Department of Biochemistry and Molecular biology, Louisiana State University Health Sciences Center, New Orleans, LA 70112,USA: Loss of α5- integrin binding domain on Nischarin leads to development of breast tumor in invivo mouse model: Altered expressions of matrix components are involved in the determination of various status of cancer progression and metastasis. In this connection, integrins are reported to function critically and they have been considered the attractive therapeutic targets in cancer research. Targeted deletion of β1-integrin in mammary epithelium do not develop breast tumors. Our previous studies explored that a novel protein Nischarin regulates breast cancer cell invasion by its efficacy in down regulating PAK, LIMK and integrin α5. It has also been revealed that cancer cell growth may be inhibited by Nischarin through limiting the expression of integrin α5. Therefore α5-integrin binding domain on Nischarin plays an important role in prevention of cancer origination. We wanted to investigate effect of loss of the integrin binding domain on Nischarin in breast cancer invivo (knock out mouse) model. Nischarin heterozygous, Nischarin null and wild type animals were used for cohort study (59 weeks) to understand the impact of Nischarin integrin binding domain in breast tumor progression. Heterozygous or Null mice were found to be affected with alopecia (hair loss) which is related to aging. To determine whether Nischarin integrin-α5 binding domain is involved in breast tumor development, we used transgenic mice carrying the mammary tumor virus-PYMT (polyoma middle T antigen) intercrossed with Nischarin heterozygous mice. PYMT mediated mammary tumor growth in mice goes through distinct morphologic stages of tumor progression comparable to progression of human breast disease and develops spontaneous metastases following disease progression. We found that reduced interaction between Nischarin and integrin-α5 expression may lead to promote breast tumor aggressiveness in mouse.

Research Affiliate Postdoc (December 2011 to September 2013) in the department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263. Single Nucleotide Polymorphisms on Aurora A kinase (SNPs). Aurora A kinase is a serine/threonine kinase, a cancer susceptibility gene because it involves in chromosome segregation and cytokinesis. Two coding single nucleotide polymorphisms in Aurora A kinase create four haplotypes. It has been reported that there is an association between these coding single nucleotide polymorphisms and esophageal cancer risk. The less kinase active Aurora A kinase haplotype combinations might induce genomic instability and increase esophageal cancer risk either in a recessive or a dominant manner. We made four haplotype of Aurora A kinase to explore proposed scientific idea in cancer biology and found that exogenous expression of the SNPs induces polyploidy. But one of the four SNPs make more colonies than other. Post-Doctoral Associate (September 2013 to September 2014), department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263. We made rat fibroblast cells exogenously expressing HRas and SNPs of Aurora A kinase to check their ability to make tumor in xenograft model. The fibroblast cells were injected subcutaneously into nude mice and the mice were monitored for tumorigecity. We found that one of four mice group delays to make tumor even all of the mice made tumor. In order to understand molecular mechanism for the differences in tumor development, the study needs to be extended further. We believe that this study can make novel signaling mechanism to understand tumor biology to get therapeutic strategy for cancer treatment.

Research Associate (September 2014 to 2016), worked in the department of Institute of Environmental Health sciences, Wayne State University, Detroit, MI. Research has been focused on cisplatin mediated toxicity which causes hearing loss. Cisplatin is used as a drug to treat cancer patients; it adversely affects to get hearing loss. The current study is focused to understand molecular mechanisms involved with cisplatin toxicity. Recently, this study has been awarded with a travel grant in 38th Midwinter Meeting “Association for Research in Otolaryngology” at Baltimore, Maryland, February 21-25, 2015. It has been reported that cisplatin mediated hair cell death is prevented by a synthetic drug SRI-110 through inhibition of protein nitration.

In 2012, pre-application was submitted for a grant (Idea award) in the department of cancer genetics, Roswell Park Cancer Institute, Buffalo, New York. Role of DNMT1 in Radiosensitivity of BRCA1 mutant breast cancer cells: In epigenetics, addition or deletion of methyl groups on the gene plays a major role in regulation of transcription, and abnormalities that lead to tumorigenesis. DNMT1, encodes a methylation maintenance enzyme, and a transcriptional target of BRCA1. In mammary tumors developed in conditional BRCA1 knockout mice, levels of DNMT1 are significantly decreased, and subsequently global DNA methylation is also reduced. Several protooncogenes, such as c-myc, c-fos and Ha-ras, are induced in those tumors, suggesting that altered gene expression due to hypomethylation of genomic DNA contributes to carcinogenesis caused by BRCA1 deficiency. From these observations, we propose our hypothesis that lowered levels of DNMT1 and DNA methylation are essential for mammary carcinogenesis in BRCA1-mutant tissues. DNMT1 is an important protein to catalyze methylation on gene, recently it was discovered as BRCA1 transcriptional target by our lab.

In 2010, a postdoctoral fellowship grant was written (submitted to DOD) in the department of biochemistry and molecular biology, Louisiana State University, LA 70112. Novel oncogenic function of Rab14: Rab (Ras-associated binding)-GTPases are members of the Ras family of small GTPases. Some of the Rab GTPases are abnormally expressed in different cancer tissues. Our data reveal that Rab14 and Nischarin interact in vitro and in vivo. Another important finding is that Rab14 is highly expressed in breast cancer tissues in contrast to Nischarin’s reduced expression in breast cancers. Furthermore, our data indicate that Nischarin reduces Rab14 driven cell invasion, possibly through PAK signaling. Thus we hypothesized that Rab14 functions as an oncogene. The intent of the proposal was to delineate the role of Rab14/Nischarin interaction in breast cancer

AWARD: Received a travel grant award in 38th Midwinter Meeting “Association for Research in Otolaryngology” at Baltimore, Maryland, February 21-25, 2015. MENTORSHIP:

Supervised postgraduate students in the department of Physiology, University of Madras, India.

(1) 2007-2008: Mr. Ramkumar, Msc., Department of Physiology, University of Madras, India. “A SIMPLE ATTEMPT BY A NEW METHOD TO REDUCE METHANOL TOXICITY - AN INVIVO STUDY”.

(2) 2006-2007: Mr. Raja, Msc., Department of Physiology, University of Madras, India. “A SIMPLE REMEDY FOR METHANOL TOXICITY”.

REVIEWER: Being a reviewer in following journals

(1) Journal Nanomicroletters

(2) Journal “Free radicals and antioxidants”

(3) Journal of biologically active products from nature

(4) Journal of basic and applied sciences- Life Science Global

(5) Pharmacognosy magazine

(6) International Journal of Pharma and Biosciences PROFESSIONAL AFFILIATIONS:

(1) Member in AAAS (American Association for the Advancement of Science)

(2) Member in AACR (American Association for Cancer Research) CERTIFICATION:

(1) Institutional Animal Care and Use Committee (IACUC) certificate

(2) Radiation safety

(3)Chemical Hygiene course

TRAINING EXPERIENCE:

(1) Participated in the Indo-UK work on designing of Doctoral Research for Ph.D. scholars held at the department of Microbiology, Dr. ALM. Post Graduate Institute of Basic Medical sciences, University of Madras on 23rd March 2005.

(2) Participated in short term course on Electrophoresis Techniques held on 6 - 11th October 2003, conducted by “ELECTROPHORESIS INSTITUTE, YERCAUD, India- 636601.

LIST OF RESEARCH PUBLICATIONS

1. Muthuvel A, Rajamani R, Senthilvelan M, Manikandan S, Sheeladevi R. Modification of allergenicity and immunogenicity of formate dehydrogenase by conjugation with linear mono methoxy poly ethylene glycol: Improvement in detoxification of formate in methanol poisoning. Clin Chim Acta, 374(1-2):122-8, (2006). 2. Arumugham Muthuvel, Rajamani R, Sundaramahalingam Manikandan, Rathinasamy Sheeladevi. Detoxification of formate by formate dehydrogenase-loaded erythrocytes and carbicarb in folate-deficient methanol-intoxicated rats. Clinica Chimica Acta, 367:162 – 169,

(2006).

3. Arumugham Muthuvel, Rajamani R, Rathinasamy Sheeladevi. Therapeutic response to single intravenous bolus administration of formate dehydrogenase in methanol-intoxicated rats. Toxicology Letters, 161: 89-95, (2006).

4. Rajamani R, Arumugham Muthuvel, Manohar Senthilvelan, Rathinasamy Sheeladevi. Oxidative stress induced by methotrexate alone and in the presence of methanol in discrete regions of the rodent brain, retina and optic nerve. Toxicology Letters, 165: 265–273, (2006). 5. Muthuvel A, Rajamani R, Sheeladevi R. Improved elimination of formate in methanol poisoning by intravenous infusion of formate dehydrogenase conjugated with linear mono methoxy poly ethylene glycol. Toxicology Letters 164, S98-S99 (2006). 6. Ramasundaram Srikumar, Narayanaperumal Jeyaparthasarathy, Sundaramahalingam Manikandan, Arumugham Muthuvel, Rajamani R. Immunomodulatory effect of Triphala during experimentally induced noise stress in albino rats. J Health Sci :53(1);142-145 (2007). 7. Rajamani R, Arumugham Muthuvel, Sundaramahalingam Manikandan, Ramasundaram Srikumar, Rathinasamy Sheeladevi.Efficacy of DL-α-LPA on methanol induced free radical changes and protein oxidative damages in folate deficient rat brain, retina and optic nerve. Chemico-Biological Interactions 167 161–167, (2007). 8. Wang Y, Rajamani R, Walch A, Alahari SK. ST14 (Suppression of Tumorigenicity 14) Gene Is a Target for miR-27b, and the Inhibitory Effect of ST14 on Cell Growth Is Independent of miR-27b Regulation. J Biol Chem. 284(34):23094-106, (2009). 9. Rajamani R, Alahari SK. Important role of integrins in the cancer biology. Cancer Metastasis Rev. (1):223-37 (2010).

10. Rathinam R, Berrier A, Alahari SK Role of Rho GTPases and their regulators in cancer progression Front Biosci.;17:2561-71 (2011).

11. Wang Y, Shenouda S, Baranwal S, Rathinam R, Jain P, Bao L, Hazari S, Dash S, Alahari SK. Integrin subunits alpha5 and alpha6 regulate cell cycle by modulating the chk1 and Rb/E2F pathways to affect breast cancer metastasis. Mol Cancer;10:84 (2011). 12. Baranwal S, Wang Y, Rathinam R, Lee J, Jin L, McGoey R, Pylayeva Y, Giancotti F, Blobe GC, Alahari SK. Molecular Characterization of the Tumor-Suppressive Function of Nischarin in Breast Cancer.J Natl Cancer Inst. (2011).13. 13. Rathinam R, Ghosh S, Neumann WL and Jamesdaniel S. Cisplatin-induced apoptosis in auditory, renal, and neuronal cells is associated with nitration and downregulation of LMO4. Cell Death Discovery;(1)-15052; (2015).

14. Jamesdaniel S, Rathinam R, Neumann WL. Targeting nitrative stress for attenuating cisplatin-induced downregulation of cochlear LIM domain only 4 and ototoxicity. Redox Biol.10:257-265; (2016).

A. Conferences & Symposia

1. Rajamani Rathinam, Samson Jamesdaniel. Presented a poster titled “LMO4 downregulation is a critical factor in cisplatin-mediated Ototoicity” at the first Brain@Wayne symposium, BrainStorm 2016, on September 29, 2016 at Wayne State University, Detroit, MI 48202.

2. Jamesdaniel S, Rathinam R. Overexpression of LMO4 mitigates Cisplatin-induced cytotoxicity in UBOC1 cells. Presented poster in Association for Research in Otolayringology, 39th midwinter meeting February 20-24, 2016. Manchester, Grant Hyatt, San Diego, California, USA.

3. Rajamani Rathinam, Samson Jamesdaniel. Cisplatin-induced cytotoxicity is associated with downregulation of LMO4 in organ of corti cell culture. Paper presented in 38th Midwinter Meeting “Association for Research in Otolaryngology” at Baltimore, Maryland, February 21-25, 2015.

4. Rathinam R, Ouchi M, Sara Cuesta Sancho, Ouchi T. Characterization of Single Nucleotide Polymorphisms of Aurora kinase A in Tumorigenesis. Abstract published at “American Association for Cancer Research” meeting at San Diego, California 2014. 5. Participated in the Roswell Park Cancer Institute Science Retreat, State University of New York (SUNY), Geneseo Campus; Jun 20-21, 2013.

6. Participated in the Roswell Park Cancer Institute Science Retreat, State University of New York (SUNY), Geneseo Campus; July 19-20, 2012.

7. Presented a poster titled “ST14 is target for miR27b and ST14 regulates tumor cell growth by upregulation of CDK inhibitor p27” in Annual research day of LSUHSC on 6th November 2009.

5. Presented a paper titled “Methanol induced free radical changes and protein oxidative damages in discrete regions of folate deficient rat brain” at the International Conference on “Toxicology, Environmental and Occupational Health”, conducted by Industrial Toxicology Research Centre, Lucknow-226001, India, held between 14th – 17th November, 2005.

6. Participated in the 1st symposium on medical Ethics & Law as a delegate conducted at Chennai, Tamilnadu, India. Sponsored by Medical Council of India, MGR Medical University and Tamilnadu medical council (19th & 20th March, 2005). 7. Presented a paper titled “Methanol induced generation of super oxide and hydroxyl radicals in rat”& Participated in the Indo-Australian Conference on Biotechnology in infectious Diseases held at Kasturba Medical College, Mahe, Manipal, Karnataka, India during 1-3 March 2005.

8. Participated in the seminar on “Recent Advancements in Physiology conducted by Department of Physiology, University of Madras on 20th February 2004 at Dr. ALM PG Institute of Basic Medical sciences, Taramani, Chennai, India. 9. Presented a paper titled “Methanol induced free radicals, an invivo study in rat” in the international conferences on “Natural products, Free Radicals and Radio protectors in Health” (NFRH-2004) held during 17th-19th Janauary, 2004, in the Department of Biochemistry, Annamalai Unversity, Annamalai Nagar, Tamilnadu, India. 10. Participated in the seminar-cum exhibition on Challenges to Biochemistry in the New Millennium conducted by Dept. of Biochemistry, Islamiah College, Vaniyambadi on Jan. 31st-Feb.2nd, 2000.

11. Participated in the satellite symposium on Free radical in Health and Diseases conducted by Dept. of Medical Biochemistry, Dr. ALM PG IBMS, University of Madras Taramani Campus, Chennai, India held on 20-22, Dec 1999.

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