Scientist » Dr. Shaji R. V

Shaji R.V 

Contact

Gene Regulation Lab,
Centre for Stem Cell Research,
Christian Medical College.

Office: +91 416 307-5114
Lab: +91 416 307-5113
Fax: +91 416 307-5103
Email: rvshaji@cmcvellore.ac.in

 

Career Interests

 

Education

  • MSc: Marine Biotechnology Goa University (1991-1993).
  • PhD: Christian Medical College, Vellore (1995-2001).
  • Post Doctoral Research Fellowship:  St Jude Children's Research Hospital, Memphis (2004-2006).
  • Post Doctoral Research Fellowship: University of Chicago, Chicago.

Awards

Research

Disease Modelling Using Induced Pluripotent Stem Cells (iPSCs):
Our laboratory is involved in disease modelling using iPSCs for understanding the molecular basis of disease pathogenesis and for drug screening on the differentiated cells derived from disease specific iPSCs. Recently, we generated iPSC lines from the fibroblasts of a patient with Fanconi anaemia (FA) by non integrative methods, after complementing the defective gene with a normal gene using a doxycycline inducible lentiviral vector. The iPSCs generated were completely characterized for their pluripotency and, in the absence of doxycycline, these cells showed the cellular phenotypes of FA including cell cycle arrest, lack of FANCD2 monoubiquitination and absence of co-localization of FANCD2 and gamma H2AX. Currently, we are using these iPSC lines to understand the molecular basis of Fanconi Anaemia disease pathogenesis. In addition to FA, using similar approach, we are creating disease models for Congenital Dyserythropoetic Anaemia (CDA) and Diamond Blackfan Anaemia (DBA).


Fig 1: Establishment of IndC-FANCA-iPSC line that exhibits the FA cellular phenotypes. (A) Schematic of molecular diagnosis of FANCA deficiency and iPSC generation. (B) Morphology of iPSCs grown on feeder layers (upper figure) and feeder-free vitronectin coated plates (lower figure). (C) Immunofluorescence analysis of NANOG, OCT4, SOX2, SSEA-4, TRA-1-81 and TRA-1-60. (D) Real-time PCR analysis of mRNA levels of NANOG, OCT4, SOX2, GDF3 AND DNMT3B, relative to the BC1 control iPSC line (data represented as mean▒SD). (E) H&E-staining of teratoma sections showing trilineage differentiation; hc-hyaline cartilage (mesoderm), ne-neuroepithelium (ectoderm) and ce-columnar epithelium (endoderm). (F) Chromosome analysis showing normal male karyotype. (G) Western blot analysis of the cells treated with Hydroxyurea showing the lack of FANCD2-monoubiquitination in the absence of DOX and FANCA expression. L and S indicate the larger monoubiquitinated and the smaller unmodified FANCD2 bands, respectively. (H) Immunofluorescence analysis showing nuclear foci formed by ?H2A.X and FANCD2. The nuclei are stained with DAPI (violet colour). In DOX+ cells, colocalization of the foci formed by ?H2A.X and FANCD2 are seen as yellow spots due to the merging of green spots of ?H2A.X foci and red spots of FANCD2 foci, and in DOX- cells, only green spots of ?H2A.X foci are seen. (I) EdU incorporation based cell cycle analysis showing a spontaneous progressive G2/M arrest in the absence of DOX and FANCA expression for two passages. (J) Alkaline phosphatase staining showing spontaneous progressive exhaustion of iPSCs in the absence of DOX and FANCA expression for two passages.

Induced pluripotent stem cells:
Introduction of four transcription factors Oct-4, Sox2, Klf4 and c-Myc into adult cells can covert them to pluripotent stem cells. These induced pluripotent stem cells (iPSCs) exhibit features characteristic of embryonic stem (ES) cells. These cells have exciting new prospects for biomedical research, drug discovery and for regenerative medicine. Our lab is interested in understanding the mechanism of the reprogramming process and the use of these cells to understand the pathophysiology of human diseases.

Generation of mouse induced pluripotent stem cells (miPSCs) from embryonic fibroblasts:

mipscs differentiation-mipscs.png

Generation of human induced pluripotent stem cells (hiPSCs) from adult dermal fibroblasts:

hiPSCs hiPSCs clones

A. miPSCs exhibited mouse embryonic stem cell like morphology and high level of expression of alkaline phosphatase (AP). Expression of other pluripotency markers SSEA-1, Oct4 and Nanog were examined by immunofluorescence.

B. miPSCs formed embryoid bodies and could be differentiated into lineages from all three germ layers. Immunostaining for (i) alpha-fetoprotein (AFP; endoderm), smooth muscle actin (SMA; mesoderm) and b3 tubulin (ectoderm) are shown.

Mechanism of globin gene regulation

During human development, three different types of hemoglobins are produced; embryonic, fetal and adult hemoglobins. In this project we are trying to understand the roles of transcription factors and epigenetics in developmental stage specific regulation of globin genes. We are also trying to understand the regulatory sequences in the globin clusters that help in stage specific expression of these genes. We are carrying out ex-vivo differentiation of haematopoietic stem cells to erythroid lineage and studying interaction of proteins with the DNA sequences using chromatin immunoprecipitation followed by microarray (ChIP-chip) to understand the roles of transcription factors and epigenetic changes in the regulation of globin genes.

Research Fellows

Kannan. V.M

Kannan. V.M

Senior Research Fellow /
PhD Student
E-mail:
kannanthoppil@cmcvellore.ac.in

Research Interests:
Generation of iPSCs for therapeutical applications.
Education:
M.Sc Biotechnology (Mahatma Gandhi University, Kerala).
Musheer Aalam

Musheer Aalam

Senior Research Fellow /
PhD Thesis Submitted
E-mail:
musheeraalam@cmcvellore.ac.in

Research Interests:
Epigenetic regulation of somatic cell reprogramming and pluripotency.
Education:
M.Sc Biotechnology (Bharathidasan University, Trichy).
Sumitha. P.B Sumitha Prameela Bharathan
Senior Research Fellow /
PhD Thesis Submitted
E-mail:
sumithapb@cmcvellore.ac.in

Research Interests:
Molecular mechanism of human somatic cells reprogramming and disease modelling of Fanconi Anemia using induced pluripotent stem cells.
Education:
M.Sc Biotechnology (Bharathiar University, Coimbatore).
Thiyagaraj. M

Thiyagaraj. M

(Appointed from Department of Haematology)

Senior Research Fellow /
PhD Thesis Submitted
E-mail:
thiyagaraj@cmcvellore.ac.in

Research Interests:
Molecular mechanism of human globin gene regulation.
Education:
M.Phil Endocrinology (University of Madras, Chennai).
Kasthuri. N

Kasthuri. N

Senior Research Fellow
E-mail:
kasthurinm02@gmail.com

Research Interests:
Education:
M.Sc Biochemistry (Thiruvalluvar University, Vellore).
Aneesha Nath

Aneesha Nath

Senior Research Fellow /
PhD Student
E-mail:
aneeshacscr@cmcvellore.ac.in
Research Interests:
Transcriptional regulation of human erythropoiesis.
Education:
M.Sc  Microbiology (VIT University, Vellore).
Abhirup Bagchi

Abhirup Bagchi

Junior Research Fellow
E-mail: abhirup@cmcvellore.ac.in
Research Interests:
Gene Therapy approaches for haemoglobin disorders.
Education:
Integrated M.Tech in Biotechnology (KIIT University, Bhubaneswar).
Smitha. I

Smitha. I

Junior Research Fellow
E-mail:
smitha.ijee@cmcvellore.ac.in

Research Interests:
Transcriptional regulation of human erythropoiesis.
Education:
M.Tech Biotechnology (Karunya University, Coimbatore).
Krittika Nandy

Krittika Nandy

Junior Research Fellow
E-mail:
krittika.nandy@cmcvellore.ac.in
Research Interests:
Disease modelling of human erythroid diseases using iPSCs.
Education:
M.E Biotechnology (BITS Pilani, Goa).
Thulaj D Meharwade

Thulaj D Meharwade

Junior Research Fellow
E-mail:
dthulaj.meharwade@cmcvellore.ac.in
Research Interests:
Disease modelling of human erythroid diseases using iPSCs.
Education:
M.Sc Microbial Gene Technology (Madurai Kamaraj University, Madurai).

Technical Staff

 

Dhavapriya. B

Dhavapriya. B

Graduate Technician
E-mail: grlab@cmcvellore.ac.in
Education:
B.Sc Microbiology (Madras University, Chennai).

Selected Publications

  • Sumitha Prameela Bharathan, Kannan Vrindavan Manian, Syed Mohammed Musheer Aalam, Dhavapriya Palani, Prashant Ajit Deshpande, Mankuzhy Damodaran Pratheesh, Alok Srivastava, Shaji RV. Systematic evaluation of markers used for the identification of human induced pluripotent stem cells. Biology Open 2017 6: 100-108; doi: 10.1242/bio.022111.
  • Srivastava A,Shaji RV. Cure for thalassemia major: from allogeneic hematopoietic stem cell transplantation to gene therapy. Haematologica December 2016 : haematol.2015.141200; Doi:10.3324/haematol.2015.141200.
  • Aalam SM, Manian KV, Bharathan SP, Mayuranathan T, Shaji RV. Identification of Stable OCT4+NANOG- State in Somatic Cell Reprogramming. Cell Reprogram. 2016 Sep 13. [Epub ahead of print] PMID: 27622636 DOI: 10.1089/cell.2016.0018.
  • Deshpande P, Kathirvel K, Alex AA, Korula A, George B, Shaji RV, Mathews V. Leukocyte Adhesion Deficiency-I: Clinical and Molecular Characterization in an Indian Population. Indian J Pediatr. 2016 Aug;83(8):799-804.
  • Kamath MS, Pradhan S, Edison ES, Shaji RV, Antonisamy B, Karthikeyan M, Mangalaraj AM, Kunjummen A, George K. Chorionic villous sampling through transvaginal ultrasound approach: A retrospective analysis of 1138 cases. J Obstet Gynaecol Res. 2016 Jun 29. doi: 10.1111/jog.13070. [Epub ahead of print]
  • Pal R, Mariappan I, Shaji RV. Editorial: Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells: Ushering of a New Era in Personalized Cell Therapies. Curr Stem Cell Res Ther. 2016;11(2):97-8.
  • Manian KV, Aalam SM, Bharathan SP, Srivastava A, Shaji RV. Understanding the Molecular Basis of Heterogeneity in Induced Pluripotent Stem Cells. Cellular Reprogramming 2015 Dec;17(6):427-40.
  • Jayasree D, Shaji RV, George B, Mathews V, Srivastava A, Edison ES. Clinical, Hematological and Molecular Analysis of Homozygous Hb E (HBB: c.79G?>?A) in the Indian Population. Hemoglobin. 2016;40(1):16-9.
  • Abraham A, Varatharajan S, Karathedath S, Philip C, Lakshmi KM, Jayavelu AK, Mohanan E, Janet NB, Srivastava VM, Shaji RV, Zhang W, Abraham A, Viswabandya A, George B, Chandy M, Srivastava A, Mathews V, Balasubramanian P. RNA expression of genes involved in cytarabine metabolism and transport predicts cytarabine response in acute myeloid leukemia. Pharmacogenomics. 2015 Jul;16(8):877-90.
  • Deshpande P, Kamalanathan N, Sampath E, George B, Shaji RV, Edison ES. Characterization of Clinical and Laboratory Profiles of the Deletional a2-Globin Gene Polyadenylation Signal Sequence (AATAAA?>?AATA-?-) in an Indian Population. Hemoglobin. 2015 Dec;39(6):415-8.
  • Mayuranathan T, Rayabaram J, Das R, Arora N, Edison ES, Chandy M, Srivastava A, Shaji RV. Identification of rare and novel deletions that cause (d▀)0-thalassaemia and hereditary persistence of foetal haemoglobin in Indian population. Eur J Haematol. 2014 Jun;92(6):514-20.
  • Mayuranathan T, Rayabaram J, Edison ES, Srivastava A, Shaji RV. A novel deletion of ▀-globin promoter causing high HbA2 in an Indian population Haematologica. 2012 Sep;97(9):1445-7.
  • David S, Jayandharan GR, Abraham A, Jacob RR, Devi GS, Patkar N, Shaji RV, Nair SC, Viswabandya A, Ahmed R, George B, Mathews V, Chandy M, Srivastava A. Molecular basis of Wiskott-Aldrich syndrome in patients from India. Eur J Haematol. 2012 Oct;89(4):356-60.
  • Varatharajan S, Abraham A, Zhang W, Shaji RV, Ahmed R, Abraham A, George B, Srivastava A, Chandy M, Mathews V, Balasubramanian P. Carbonyl reductase 1 expression influences daunorubicin metabolism in acute myeloid leukemia. Eur J Clin Pharmacol. 2012 Dec;68(12):1577-86.
  • Jain S, Edison ES, Mathews V, Shaji RV. A novel d-globin gene mutation (HBD: c.323G>A) masking the diagnosis of ▀-thalassemia: a first report from India. Int J Hematol. 2012 May;95(5):570-2.
  • Edison ES, Sathya M, Rajkumar SV, Nair SC, Srivastava A, Shaji RV. A novel ▀-globin gene mutation HBB.c.22 G>C produces a hemoglobin variant (Hb Vellore) mimicking HbS in HPLC. Int J Lab Hematol. 2012 Oct;34(5):556-8.
  • Abraham A, Varatharajan S, Abbas S, Zhang W, Shaji RV, Ahmed R, Abraham A, George B, Srivastava A, Chandy M, Mathews V, Balasubramanian P. Cytidine deaminase genetic variants influence RNA expression and cytarabine cytotoxicity in acute myeloid leukemia. Pharmacogenomics. 2012 Feb;13(3):269-82.
  • Edison ES, Venkatesan RS, Govindanattar SD, George B, Shaji RV. A novel 26 bp deletion [HBB: c.20_45del26bp] in exon 1 of the ▀-globin gene causing ▀-thalassemia major. Hemoglobin.2012;36(1):98-102.
  • Edison ES, Shaji RV, Chandy M, Srivastava A. Interaction of hemoglobin E with other abnormal hemoglobins. Acta Haematol. 2011;126(4):246-8.
  • Mathews V, George B, Chendamarai E, Lakshmi KM, Desire S, Balasubramanian P, Viswabandya A, Thirugnanam R, Abraham A, Shaji RV, Srivastava A, Chandy M. Single-agent arsenic trioxide in the treatment of newly diagnosed acute promyelocytic leukemia: long-term follow-up data. J Clin Oncol. 2010 Aug 20;28(24):3866-71.

  • Complete list of publications

    https://www.ncbi.nlm.nih.gov/pubmed/?term=shaji%20RV%5BAuthor%5D%20or%20Velayudhan%20SR%5BAuthor%5D&cmd=DetailsSearch

Pre-clinical lentiviral gene therapy

Dr. Alok Srivastava, Centre for Stem Cell Research, CMC, Vellore.

Disease modelling of haematological disorders using iPSCs

Dr. Alok Srivastava, Centre for Stem Cell Research, CMC, Vellore.
Dr. Biju George, Department of Haematology, CMC, Vellore.

Identification of genetic and epigenetic factors involved in stem cell maintenance and differentiation using RNA interference

Dr. Sailu Yellaboina, CR Rao Advanced Institute of Mathematics, Statistics & Computer Science, Hyderabad.
Dr. Sreenivas Kuruketi, University of Hyderabad.
Dr. Sanjay Kumar, Centre for Stem Cell Research, CMC, Vellore.
Dr. B. Poonkuzhali, Department of Haematology, CMC, Vellore.


Bioinformatics

Dr. Prashant Kumar, Institute of Bioinformatics, Bengaluru.

 

  • Janakiram Rayabaram
    PhD Student (2009-2015)
    Current position: Scientist - I, Aurigene Discovery Technologies Limited, Bangalore.
  • Nancy Beryl Janette
    PhD Student (2009-2015)
    Current position: Lecturer, Department of Haematology, Christian Medical College, Vellore.

"Content will be uploaded soon"