Page 84 - Annual report 2021-22

Annual Report 2021-22 |

Suz12 siRNA using a peptide M9 and Chondroitin sulphate (CS) as delivery agents, there was an almost
25 % decrease in cellular transcript levels of Suz12 and caused a significant visible increase in wound
closure in a scratch assay in HaCaT cells. miR-497 mimic transfection using the M9 and CS significantly
increased cellular miR-497 levels and a modest increase of rate of wound closure was observed.
Conversely when the same delivery agents were used to transfect the miR-497 inhibitor into cells, a
time dependent inhibition in wound closure was observed in a scratch assay in HaCaT cells. Initial 67
results show that overexpression of Dicer in keratinocyte cells promote migration/proliferation and
significant closure of the wound area in the Dicer transfected cells as compared to control cells. The
transfection experiments were done in collaboration with Munia Ganguli.

Malabika Datta's groups also explored the epigenetic consequences of miR-449a, miR-539 and lncRNA
H19 in the skeletal muscle of animal models of obesity and diabetes. In vitro and in vivo mechanistic
validation of the identified ncRNA mediated epigenetic deregulation and consequent roles in aberrant
cell metabolism are being pursued.

miR-359-5p levels are decreased and DNMT3b levels are up-regulated in the skeletal muscle in db/db
and high-fat diet fed mice. Using validation protocols, miR-539-5p was validated to target DNMT-3b.
Mechanistically, miR-539-5p-DNMT-3B axis was found to alter levels of Srebf1 in the skeletal muscle.
In vivo antagonism of this miRNA in normal mice induced hyperglycemia and impaired oral glucose
tolerance. Another ncRNA, lncRNA H19 that is significantly decreased in the skeletal muscle during
diabetes, elevates HDAC6 levels and this H19-HDAC6 regulation impairs insulin signaling by down-
regulating IRS1 levels in the skeletal muscle.

Again, I see no reason why the belief that we are insignificant or
fortuitous should lessen our faith.” — Rosalind Franklin