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CURRICULUM VITAE
KAVITHA GOWRISHANKAR
Postdoctoral research fellow, Melanoma Research, Hormones and Cancer
University of Sydney at Kolling Institute of Medical Research, L8
Royal North Shore Hospital, St. Leonards, Sydney, NSW 2065
Tel: 61-414944648
[email protected]
Currently a postdoctoral fellow dissecting the mechanisms of acquired resistance to
targeted therapy in melanoma and identifying novel targets to improve treatment
options.
Technical skills:
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Tissue culture: mammalian cell culture, transient transfections, creating stable
cell lines, establishing primary cell lines from patient tumors; Transfections
and Lentiviral transductions of primary cells and cell lines
Molecular and cell biological techniques including flow cytometry, viability
assays, Western blotting, PCR, RT-PCR, Q-PCR, cloning
Immunoprecipitation, TUNEL (on cells and tissue sections), ELISA assays
Handling of virus, Infectious centre assays for viral titre determination
Culturing cell free virus, cell associated virus
Handling and Dissection of human fetal tissue and tumors
Immunohistochemistry, immunofluorescence staining, In-situ hybridisation
Handling, breeding and maintenance of transgenic mice and analysis
Good written skills; fluent in oral communication and presentations,
manuscript preparation
Design of experiments and research proposals, implementing projects and
guiding assistants
Tutoring and training students (trained two Hons students 2007 and 2005) Cosupervisor for a PhD student (2011- current)
Work experience:
Time
2013current
Position
Postdoctoral
fellow
Place
Kolling
Institute of
Medical
research,
Project
PD-L1
regulation in
melanoma cells;
improving
Expertise
Extensive cell culture,
preclinical models,
drug interaction, cell
biological assays
20092013
Postdoctoral
fellow
20082005
PhD
20052003
Research
Assistant
20032001
Research
Assistant
19981996
Research
Assistant
Royal North
Shore
Hospital,
University of
Sydney
immunotherapy
Westmead
Millennium
Institute
(WMI),
Westmead
Institute for
Cancer
research,
University of
Sydney
Centre for
Virus
research,
WMI,
University of
Sydney
Resistance to
MAPK
inhibition in
MELANOMA
Centre for
Virus
research,
WMI,
University of
Sydney
Dept. of
Pathology,
University of
Michigan,
USA
RIKEN,
Tsukuba Life
Sciences
Centre
Varicella zoster
virus infection
of human
ganglia
Varicella zoster
virus infection
of human
ganglia
Characterization
of NRADD, a
protein involved
in neuronal
apoptosis
Characterization
of transgenic
mice expressing
HIV receptors
including flow
cytometry, western
blotting, primary cell
lines; currently
establishing T-cell
cultures and CTL
assays
Extensive cell culture,
preclinical models,
drug interaction, cell
biological assays
including flow
cytometry, primary cell
lines
Immunohistochemistry,
Immunofluorescence,
Cell culture, human
explant ganglia,
ELISAAlso involved in
tutoring, training and
teaching Hons students
and assistants
Immunohistochemistry,
Immunofluorescence,
Cell culture, human
explant ganglia, ELISA
Cell culture, western
blotting, molecular
techniques like cloning
Transgenic mice, Flow
cytometry, invasion
assays
Academic Background:
Degree/Position
Institution
PhD
University of
Sydney
Indian Institute
Research training
Years
studied/worked
3.5 years
3 years
Dates
Grades/marks
20052008
1992-
CGPA 7.2
Master of ScienceBio-Medical Genetics
Bachelor of ScienceBotany
Schooling
of Science,
India
University of
Madras, India
University of
Madras, India
India
1995
2 years
3 years
12 years
19901992
19871990
19751987
77%
81.9%
85.92%
Distinctions:
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APA scholarship from March 2005 for three and a half years
CSIR (Council for Scientific and Industrial research), India, research fellow
for the period Sept 1992-Nov 1995
First rank holder (gold medallist) in the University of Madras, India, for
Master of Science (Bio-medical Genetics) 1992
First Rank holder (gold medallist) in the University of Madras, India, for
Bachelor of Science (Botany) 1990
Details of work done
Postdoctoral training from May 2013 to current
In continuance with training in melanoma biology, the current project involves
studying signalling pathways in melanoma cells to identify novel targets to increase
the efficacy of current treatments. Immunotherapy has shown remarkable response in
the treatment of melanoma with Ipilumumab obtaining FDA approval. Recent clinical
trials have shown promising success with anti-PD1 and anti-PD-L1 antibodies being
used in treatment. The laboratory’s focus is using epigenetic modulators to increase
apoptosis of melanoma cells. To fit in with that focus, my specific project aims to
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Study epigenetic regulation of PD-L1 in melanoma cells and those with
acquired resistance to targeted therapy (BRAF and MEK inhibitor)
Establish primary cell lines from patient tumors and study epigenetic
regulation of key pathways and proteins involved in resistance and apoptosis
Establish both allogeneic and autologous CTL assays
The overall aim of the project is to improve targeted therapy with a focus on
improving immune function
Postdoctoral training from 2009 to April 2013
Melanoma is a highly aggressive form of cancer which is refractory to conventional
chemotherapy. The advent of targeted therapy, using mutant BRAF specific inhibitors
has revolutionised treatment. Even though there is remarkable response, patients
relapse within a short period of time. The project involved developing preclinical
models of acquired resistance. Understanding the mechanisms of resistance can help
design novel rational combinations of therapies to combat metastatic melanoma.
Specific contribution involved the development of pre-clinical models of BRAF
inhibitor resistance, using melanoma cells and conducting detailed cell biological
assays. Results were instrumental in obtaining a successful NHMRC project funding
for the lab and a program grant as part of the melanoma institute of Australia;
Establishing primary cell lines (ex vivo) from metastatic tumor (patient derived cell
lines being a step towards personalised medicine) and training students and research
assistants in the lab.
Techniques involved:
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Cell culture- maintenance, establishing drug resistant clones, transfection and
transduction, treatment with drugs and small molecule inhibitors
Analysing Response to drugs /inhibitors
o Viability assays (MTT)
o Cell cycle assays by Flow Cytometry
o Apoptosis- Annexin V staining analysed by Flow Cytometry
o Western blotting for signalling pathway activation,
o Tumor tissue handling and establishing ex vivo cell lines
o Microarrays for gene expression (collaboration with colleagues)
o Kinase arrays for studying activated signalling proteins
o Next generation exome sequencing (Axeq).
(Jan 2009-2013- One first author publication, one second and one thrid author
publication. A few others as part of other collaborative projects in the lab).
PhD following research assistant position at Centre for Virus Research, WMI,
University of Sydney 2003-2008
The Abendroth laboratory is interested in studying the immunopathology of Varicella
Zoster Virus (VZV), the causal agent for chicken pox in humans.
PhD thesis: In continuation with the research project (as an assistant) in the VZV lab,
extensive immuno-histochemical analyses on human ganglia collected after the
incidence of herpes zoster, showing reactivation of Varicella Zoster Virus, were
carried out. Novel findings of the nature of immune infiltrate, were reported during
oral presentations at the International Herpes Workshop (IHW, Seattle 2006) and
published later. An in vitro explant ganglia model using human fetal tissue was
established (published in J. Virol 2007) and continues to be used by other students to
study VZV infection. The infected explanted ganglia were shown to secrete key IFN
induced chemokines. These studies have contributed valuable information on VZV
pathogenesis in neural cells that will aid in developing better immune based targeted
therapies to combat herpes zoster, and its painful complication, post herpetic
neuralgia (PHN). (Duration 3.5 yrs- three publications)
The establishment of protocols and generation of results paved the way for 3 Honours
projects and two PhD projects. As a research assistant, prior to registering for the
doctoral program, I was primarily responsible for setting up the IHC expertise in the
lab, in addition to various other roles.
Techniques involved:
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Immunohistochemistry and immunofluorescence
Genomic, RT, Q- PCR, In-situ hybridisation
Detection of apoptosis in the paraffin sections of zoster ganglia-by TUNEL
Establishing an in vitro explant ganglia model using fetal ganglionic tissue
ELISA assays, Q-PCR
Research at the University of Michigan, Ann Arbor, USA – 2001-2003
Synopsis of research: Analysis of NRADD, (neurotrophin alike death domain
protein), a novel transmembrane death domain containing protein (that was
discovered in the lab) that is similar to p75 NTR in the C-terminus but has an unique
N-terminal ectodomain. NRADD causes endoplasmic reticulum (ER) mediated
apoptosis in certain neuronal cells. It also interacts with the ER resident proteins of
the gamma-secretase complex, is processed by the  -secretase and metalloproteases
and affects the generation of amyloid protein products.
(Duration – 1.5 years with Dr Claudius Vincenz – Two publications)
Techniques involved:
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Cloning of rat NRADD by RT-PCR from rat striatal neuronal cells.
Creating stable/inducible cell lines of the different mouse NRADD mutants
and carried out death assays of these mutants.
Protein purification for use as an antigen to raise monoclonal antibodies. All
preliminary screenings were done by ELISA and ascites checked on Westerns.
Western Blotting/ Immunoprecipitations to detect proteins in cell lysates. in
vitro de-glycosylation and extensive transient transfection studies in various
cell lines to characterize the glycosylation and proteolytic processing patterns
of the different NRADD forms.
Carried out experiments that induce -secretase and metalloprotease mediated
processing which are blocked by specific inhibitors.
Research at RIKEN, Tsukuba Life Sciences Centre, Japan- 1996-1998
Synopsis of research: Worked as part of the team that developed a mouse model
system for studying AIDS pathogenesis. Transgenic mice expressing hCD4 and
hCXCR4 were made and analysed in detail. Thymocytes from these mice were
infected with T-tropic HIV. Tissue distribution of CD4+CXCR4+ cells were studied
and it was found that there was severe loss of circulating peripheral blood CD4 cells
in the transgenics (similar to AIDS patients) which preferentially migrate to other
issues like the bone marrow, mainly due to over expression of CXCR4. Blocking Gprotein expression by treating with pertusis toxin reversed the effect as assessed by in
vivo cell migration assays.
(Duration 1.5 years with Dr Shinichiro Sawada- One publication)
Techniques involved:
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Was responsible for screening, maintaining, breeding the transgenic mice.
Analyzed the transgenic tissues by Flow Cytometry (FACS).
Performed all the in vivo cell migration assays.
Undertook basic training in cell culture, monoclonal Ab production and
standard molecular techniques and helped in the maintenance of cell lines.
Research training at the Indian Institute of Science (IISc), India
Joined the IISc as a junior research fellow at the Department of Microbiology and
Cell Biology. Undertook courses in Microbiology, advanced Molecular Biology,
principles of genetic Engineering, Virology and basic laboratory techniques.
Abstract of Work: Transcriptional regulation of fibroin gene in Bombyx mori was
studied as an example of differential gene expression in eukaryotes.
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Towards cloning the full length fibroin gene, genomic DNA isolated from the
larval instars was probed with radiolabeled sequences-Southern Blotting.
Subcloning of the 1.4 kb upstream regulatory sequences were carried out to
facilitate DNA-Protein interaction studies. Crude and Heparin sepharose
fractionations of nuclear extracts were used in standard gel retardation
assays to obtain specific complexes.
Differential display PCRs were performed to study other messages to compare
with fibroin.
Publications:
1. Acquired resistance to BRAF inhibition can confer cross-resistance to
combined BRAF/MEK inhibition- Gowrishankar K, Snoyman S, Pupo,
GM, Becker TM, Kefford RF, Rizos, H, J. Invest Dermatol 2012 Jul;
132(7):1850-9
2. Carlino, M. S., K. Gowrishankar, et al. (2013). "Antiproliferative
effects of continued mitogen-activated protein kinase pathway
inhibition following acquired resistance to BRAF and/or MEK
inhibition in melanoma." Mol Cancer Ther 12(7): 1332-1342
3. Carlino, M. S., J. R. Todd, Gowrishankar K, et al. "Differential activity
of MEK and ERK inhibitors in BRAF inhibitor resistant melanoma."
Molecular Oncology 2014 May:8 (3)544-54
4. Control of NF-kB activity in human melanoma by bromodomain and extraterminal protein inhibitor IBET-151- Gallagher SJ, Mijatov B, Gunatilake D,
Gowrishankar K, Tiffen J et al Pigment Cell Melanoma Res 2014 Jun 13
(Epub ahead of print)
5. The epigenetic regulator IBET-151 induces BIM-dependent apoptosis and
cell cycle arrest of human melanoma cells-Gallagher SJ, Mijatov B,
Gunatilake D, Tiffen J, Gowrishankar K et al-J Invest Dermatol 2014 Jun 6
(Epub ahead of print)
6. “Acquired resistance to targeted MAPK inhibition in melanoma” Chapter
7 in “Melanoma - From Early Detection to Treatment", Kavitha
Gowrishankar, Matteo Carlino, Helen Rizos; Jan 30 2013 under CC BY
3.0 license DOI:10.57772/50853 ISBN 978-953-51-0961-7, hard cover,
p719, Publisher- InTech, Ed. Guy Huynh Thien Duc. Book Chapter
7. Anti-proliferative effects of continued mitogen activated protein kinase
pathway inhibition following acquired resistance to BRAF and/orMEK
inhibition in melanoma –M.S. Carlino, Kavitha Gowrishankar et al.
Mol. Cancer Ther. 2013 Jul; 12(7):1332-42
8. Absence of distinguishing senescence traits in human melanocytic
nevi- Tran SL, Haferkamp S. Scurr LL, Gowrishankar K et al, J. Invest
Dermatol 2012 Sep:132(9):2226-34
9. Oncogenic B-RAFV600E signaling induces the T-box3 transcriptional
repressor to repress E-cadherin and enhance melanoma cell invasionS.C Boyd, B.Mijatov, G.M.Pupo, S.Tran, K.Gowrishankar, H.M.
Shaw, C.R Goding, R.A.Scolyer, R.F.Kefford, H.Rizos, T.M.Becker,
J.Invest.Dermatol 2012. J Invest Dermatol. 2013 May;133(5):1269-77
2012 Nov 29. doi: 10.1038/jid.2012.421. [Epub ahead of print]
10. Mutant BRAF-Mcl1 survival signalling depends on the STAT3
transcription factor Oncogene 2012 T.M. Becker, S.C.Boyd, Mijatov
B, K.Gowrishankar et al 2013, Oncogene mar 4 doi:
10.1038/onc.2013.45 [Epub ahead of print] 4 March 2013;
doi:10.1038/onc.2013.45.
11. Upregulation of CXCL10 in human dorsal root ganglia during
experimental and natural varicella-zoster virus infection- Steain M,
Gowrishankar K, Rodriguez M, Slobedman B, Abendroth A, J. Virol
2011 Jan:85(1):626-31
12. Characterization of the host immune response in human ganglia after
herpes zoster-Gowrishankar K, Steain M, Cunningham AL, Rodriguez
M, Blumbergs P, Slobedman B, Abendroth A. J. Virol 2010
Sep:84(17):8861-70
13. Productive varicella-zoster virus infection of cultured intact human
ganglia-Gowrishankar K, Slobedman B, Cunningham AL, MirandaSaksena M, Boadle RA, Abendroth A; J. Virol 2007 vol 81(12):6752-6
14. Release of a membrane bound death domain by -secretase processing
of the p75 NTR homologue, NRADD-Kavitha Gowrishankar, Michael
Zeidler and Claudius Vincenz; J. Cell. Sci. 2004 117: 4099-4111.
15. NRADD, a novel membrane protein with a death domain involved in
mediating apoptosis in response to ER stress- Wang X, Shao Z,
Zetoune F, Zeidler M, Kavitha Gowrishankar and Vincenz C; Cell
Death and Differentiation 2003 vol 10, p580-591.
16. Disturbed CD4+ T cell homeostasis and in vitro HIV-1 susceptibility
in transgenic mice expressing T-cell line tropic receptors-S. Sawada,
Kavitha Gowrishankar, R. Kitamura et al. 1998 J. Exp. Med, 187, No:
9 p 1439-49.
Presentations:
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AACR, San Diego 2014 Oral presentation at the minisymposium –Gene
regulation and transcription factors “Regulation of PD-L1 expression in
human melanoma by NF-kB”
Lorne Cancer Conference 2014, Lorne, Vic, Australia “ Regulation of PD-L1
expression in human melanoma by NF-kB” poster presentation
Oral presentation “Dual MAPK inhibition in melanoma is not driven by
known mechanisms of resistance” SCC2012, Sydney Cancer Conference,
University of Sydney, Sept 2012
Poster presentations and abstracts submitted at the international conference of
the Society of Melanoma Research (SMR) 2010, 2011 and 2012
Two Oral presentations at the IHW (International Herpes Workshop, Seattle,
USA, 2006)
Poster presentation at Australasian Society of Immunology (ASI) conference,
Sydney, 2007
Hospital week, Westmead Hospitals and WSAH annual science week, 2004,
2005, 2006
Poster presentation at National Virology Group conferences 2005
References:
1. Prof. Peter Hersey, MD, D.Phil, FRACP, Melanoma Research, Level
8, Kolling Institute of Medical Research, Royal North Shore Hospital,
The University of Sydney, St. Leonards, NSW 2065, Australia – Tel :
+61-2-99264715 Fax: +61-2-99264034 Email:
[email protected]
2. Assoc. Prof Georgina Long MBBS, PhD, FRACP, Melanoma Biology
and Translational Research, Melanoma Institute of Australia, The
University of Sydney, 40, Rocklands Rd, North Sydney, NSW 2060
Tel: +61-2-99117200 Fax :+61-2-9954 9418 Email:
[email protected]
3. Assoc. Prof. Allison Abendroth PhD, (Immunology and Infectious
Diseases) Central Clinical School, D06, Blackburn Building, The
University of Sydney, NSW 2006 Australia. Tel: +61-2-9351 6867
Fax: 61-2-9351 4731 Email: [email protected]
4. Prof. Tony Cunningham, PhD Director, Westmead Millennium
Institute, Westmead, and The University of Sydney, Faculty of
Medicine, PO box 412, NSW 2145 Australia. Tel: +61-2-9845 9100
Fax: 61-2-9845 9005 Email:[email protected]
5. Prof. Richard Kefford, Professor of Cancer Medicine, Macquarie
University, Co-Director of Research and Melanoma Institute of
Australia 40, Rocklands Rd, North Sydney, NSW 2060 Tel: +61-299117200 Fax :+61-2-9954 9418
Email:[email protected]
Other information:
Date of Birth: 4th August 1970
Nationality: Australian citizen and OCI (Overseas Citizen of India)
Sex: Female
Break in career from 1998 to end of 2001 to start a family.