The GBMD is interested in understanding the role of transcription factors and small RNAs in health and disease.  In particular, we are interested in understanding the role of tumor suppressor p53–the most mutated gene in human cancer–and its structural and functional homologues p63 and p73 in Cancer, Diabetes, Myocardial disorders, ageing, Hypertension, Memory, learning, longevity, reprogramming, Stem cell biology, regenerative medicine and neurodegenerative diseases.  p53, p63 and p73 are transcription factors. They respond to variety of stresses to protect the genome and therefore they have generally been referred as “Guardians of the genome” (Boominathan, 2010, Cancer and Metastasis review).

a. Identifying the networks of tumor suppressor genes & understanding how they are regulated:

Bioinformatics tools, computational/comparative genomics tools, and the knowledge-based system biology approach will be used to identify:

a) tumor suppressor networks; and

b) tissue specific tumor suppressor pathways.  Then, the identified tumor suppressor networks/pathways will be characterized biochemically and genetically.

b. Molecular mechanisms of tumor suppression:

Bio-chemical experiments that test the role of

(i) the tumor suppressor p53/p63/p73 in the regulation of miRNAs and its target gene networks;

(ii) TA-p63, TA-p73 and p53 in the regulation of self-renewal;

(iii) the tumor suppressors TA-p63, TA-p73 and p53 in the inhibition of Cancer stem cell generation; and

(iii) TA-p63, TA-p73 and p53 in the regulation of Epithelial to mesenchymal transition, invasion, migration, and metastasis, will be carried out.

c. Role of p53/p63/p73 in the regulation of self-renewal and reprogramming: 

Bio-chemical experiments that test the role of

(i) p53, p73, and p63-related proteins in the regulation of reprogramming factors;

(ii) DNp63 in the regulation of stem cell renewal;

(iii) p53, p63 and p73 in the regulation of miRNA processing enzymes

(iv) p53, p63 and p73-dependent target genes in the regulation of stem cell-renewal gene network; and

(v) p63/p73-DNMT axis in stem cell renewal and reprogramming, will be carried out.

d. Identification of agents that promote reprogramming of differentiated patient-specific cells into pluripotent stem cells:

A number of tumor suppressors appear to function as a bottleneck against cellular reprogramming into induced pluripotent stem cells.  Generation of induced pluripotent stem cells is of great importance in degenerative diseases (eg., Diabetes, Parkinson’s and Heart attack).  Therefore, identification of agents that:

(i) transiently down regulate the expression of tumor suppressors and their target genes; and

(ii) increase the expression of reprogramming factors in differentiated cells will be useful in generating induced pluripotent stem cells.  These pluripotent stem cells can be used to obtain desired cell type to treat patients with degenerative diseases.

e. Understanding p63/p73/p53 regulated molecular processes and its associated disease conditions:

 Identifying the activators or suppressors of transcription factors and p53-related proteins that will be useful in

i) curing cancer, Diabetes, Hypertension, & Myocardial disorders;

ii) eradicating microbial infections/tuberculosis;

iii) promoting microbial resistance and immunity

iv) increasing stress resistance

v) suppressing ageing process &

vi) increasing longevity

Genetic screens in mammalian cells will be carried out to identify small molecules that increase or decrease the expression of transcription factors/tumor suppressor genes and their target genes.  Then, the identified activators or suppressors will be characterized  biochemically to test whether they:

(i) confer resistance against several bacterial and viral infections;

(ii) cure cancer, diabetes, hypertension and myocardial infarction; and

(iii) improve learning, longevity, memory, & ageing .

f. Understanding the role of p53, p63 and p73 in the  regulation of small RNAs processing:

p53, p63, and p73 had been shown to function as both positive and negative regulators of the miRNA processing components (Boominathan, Plos One, 2010).  Further, we intend to characterize how p53, p63, and p73 function as regulators of miRNA/piRNA/siRNA processing.

g. Targeting tumor suppressors through miRNAs in human cancer:

Bio-chemical experiments that test the ideas such as:

(i) increasing the stability or expression of tumor suppressor genes in human cancer through miRNAs;

(ii) targeting multiple tumor suppressors simultaneously through miRNAs/protein degrading components of proteasomes ;

(iii) identifying small molecules that increase the expression or stability of multiple tumor suppressors simultaneously in human cancer; and

(iv) increasing the expression of p53, p63, and p73 and their target miRNAs to inhibit Epithelial to mesenchymal transition, metastasis, invasion, and cancer stem cell proliferation, will be carried out.

h. Molecular Therapeutics:

a) Genetic screening:

Small molecule or Bio-molecule screening will be conducted in Mammalian cells using multiple-promoter containing expression vectors for the chosen therapeutic targets.

b) miRNA therapeutics:

miRNAs-based therapeutic program will be initiated for a number of targets identified and characterized at the GBMD.  This will be useful in curing cancer, Diabetes, Hypertension, & Myocardial disorders and increasing stress resistance/longevity.

c) Regenerative/Stem cell therapeutics:

miRNAs-based stem cell therapeutic program will be initiated for targets identified at the GBMD.  Genetic screens in mammalian cells will also be setup to identify small molecules or Bio-molecules that activate cellular targets with regenerative property.

d) Mutation analysis/diagnostic test:

i) tumor suppressors identified from the tumor suppressor network analysis will be screened for mutations;

ii) expression of therapeutic targets chosen will be determined in patient samples; and

iii) diagnostic/prognostic tests will be developed in coordination with Clinicians/Physicians.

 Copy right protected by Genome-2-Bio-Medicine Discovery Center (GBMD) 9/November/2012