Module 2: Novel Approaches for the Generation of Clinically Relevant iPS Cell Lines

Mouse induced pluripotent stem cells expressing an Oct4-GFP transgene.

Module Leader Dr Paul J. Verma
Host Organisation Monash Institute of Medical Research, Monash University

Module description

Reprogramming using an iPS (induced pluripotent stem cell) approach appears to share similarities with other reprogramming approaches such as SCNT and cell fusion, however there are subtle differences in the timing and extent of reprogramming achieved by each approach.

This module believes that while this research is incredibly exciting and promising, several issues need to be addressed for iPS technology to be clinically relevant.

The laboratory has optimised generation and characterisation of iPS cells from human and mouse somatic cells. Preliminary studies in the laboratory have identified approaches to significantly increase the efficiency of iPS cell generation; they include altering epigenotype of the somatic cells and/or selecting specific cell types as iPS targets. The laboratory have also developed an approach to efficiently deliver proteins in a sustained manner to the nucleus of somatic cells and are examining the generation of iPS cells following the direct delivery of specific proteins.

The overall aim of the project is therefore to investigate approaches to efficiently generate clinically relevant human iPS cells and lines of disease. And further, to disseminate this information and material to the Australian research community.

The image to the right was created via somatic cells from an Oct4-GFP transgenic mouse line that were reprogrammed to pluripotency by forced expression of Oct4, Sox2, cMyc and Klf4 genes. This has resulted in re-activation of the Oct4-GFP transgene integrated in the somatic genome.

Aims

  1. Compare various cell types from different lineages to effect efficient and optimal generation of iPS cells: 'Cell types from different lineages will reprogram at different efficiencies'.
  2. Investigate approaches to generate iPS cells without genetic modification, using non-integrating viruses and protein delivery to somatic cell nuclei. 'Delivery of key Pluripotency genes or proteins via rAAV or proteins, respectively, to somatic cells will induce Pluripotency without genetic modification of the cells'.
  3. Effect efficient generation of human iPS cells 'Approaches developed to generate iPS cells from murine tissues can be translated to human cell lines'.

Module Leader biography

Dr Paul Verma obtained his PhD at Adelaide University and subsequently joined BresaGen Ltd, Adelaide in 1995, working on transgenesis and nuclear transfer aimed at developing pigs with organs suitable for xeno-transplantation. He later joined the Luminis-BresaGen Cell Therapy Program, where he established a research program aimed at providing alternatives to therapeutic cloning.

In 2001 he moved to the Monash Institute of Medical Research, where he currently heads the stem cell program. His research aims to produce autologous (patient-specific) stem cells, using cell-reprogramming techniques such as somatic cell nuclear transfer and iPS cell technology that are clinically relevant and transplantable. He serves on the NHMRCs, Cellular Therapies Advisory Committee (CTAC), on the editorial board of Animal Reproduction Science and is a visiting Professor at the National Centre for Biological Sciences, Bangalore, India.

Contact details

 E-mail  paul.verma@med.monash.edu.au
 Phone   +61 3 9594 7000
 Web
  www.monashinstitute.org

Selected publications

  1. Sumer H, Tat PA, Jones KJ, Liu J and Verma PJ. (2009) JOURNAL OF STEM CELLS. In press. (Invited review).
  2. Tat P, Sumer H, Jones, K and Verma PJ (2009) In“Stem Cells: Basics and Applications“. Ed Deb K. McGraw Hill (In press)
  3. Sumer H, Jones KJ, Liu J, Rollo BN, van Boxtel AL, Pralong D and Verma PJ. (2009) STEM CELLS AND DEVELOPMENT; Feb 17. [Epub ahead of print]
  4. Liu J, Jones KL, Sumer H and Verma PJ. (2009) MOLECULAR REPRODUCTION AND DEVELOPMENT; 76(6):580-6
  5. Bhartiya D, Ritchie W, Hinduja I, Nandedka P, Zaveri K, Meherji P, Mukadam L, ChohanP, Patwardhan A, Nagvenkar P, Kumar N, Tat P, Richings NM and Verma PJ 2008 JOURNAL OF STEM CELLS; 3 (4): 255-64
  6. Verma PJ & Trounson AT, Editors - Methods in Molecular Biology “Cell Reprogramming and Transgenesis by Nuclear Transfer”. Humana Press, NY. 2006.
  7. Pralong, D; Trounson, AO; Verma, PJ 2006 STEM CELL REVIEWS 2 (4):331-340
  8. Pralong D, Mrozik K, Occhiodoro F and Verma PJ. (2006) Reprogramming differentiated nuclei with pluripotent cytoplasm In: Cell Reprogramming and Transgenesis by Nuclear Transfer. Eds: Verma, P.J. and Trounson, A.O. Humana Press, NY, USA
  9. Pralong, D; Mrozik, K; Occhiodoro, F; Wijesundara, N; Sumer, H; Van Boxtel, AL; Trounson, A; Verma, PJ 2005 CLONING AND STEM CELLS 7 (4):265-271
  10. Pralong, D; Lim, ML; Vassiliev, I; Mrozik, K; Wijesundara, N; Rathjen, P; Verma, PJ 2005 2005 CLONING AND STEM CELLS 7 (4):272-278