Identification And Characterization Of Somatic Mutations Associated With Progression Of Acute Myeloid Leukemia Having Flt3 Itd

691 words - 3 pages

My thesis dissertation titled “Identification and characterization of somatic mutations associated with progression of Acute Myeloid Leukemia having FLT3ITD and screening of small-molecule inhibitors in treatment of AMLFLT3ITD” at Cancer Science Institute (CSI) of Singapore, National University of Singapore (NUS) was the ideal experience to acquire skills in molecular cloning, cell culture, protein biochemistry, bioinformatics and exome sequencing.

The molecular pathogenesis of Acute Myeloid Leukemia has not been completely deciphered yet but develops through a multistep acquisition of a wide range of somatic mutations. Therefore, investigation of cancer genome sequences and structures provide insights for understanding cancer biology, diagnosis and therapy. Recent progress in understanding the biology of this disease and identification of driver mutations has ushered in a new era of molecular therapeutics. In this study, we observed several such somatic mutations and the role of these mutations in the progression of the disease. We performed whole exome sequenced on 11 cases of AMLFLT3ITD from diagnosis (Dx) complete remission (Cr) and relapse (Rel). We validated approximately 10-12 mutations per patient. We validated total 137 mutations in 132 genes from 11 cases by Sanger sequencing. These 137 mutations include single nucleotide variation (SNVs), small insertions and deletions (Indels). 60 mutations were present only in Dx and 20 were validated as Rel specific. In addition to this we observed 52 were present in both Dx and Rel. We identified and verified mutations in FAT1 in FLT3ITD subtype and studied the biology of these mutations in AML cells having FAT1 knockdown and overexpression resulting in a tumor suppressor-like activity. We are studying biology of mutations in MLL3 likewise.
Another part of this project was screening for small-molecule inhibitors in the treatment of AML FLT3ITD subtype. I screened set of 100 compounds and studied the effect of each on AML cell lines and traced signaling mechanism in few. Both these projects are in preparation for publication.

During this period, I was involved in two other projects on biology of Lamc2 in tumor progression of Anaplastic thyroid cancer and Pancreatic Cancer. The Thyroid Cancer study titled “Laminin-5 gamma-2 (LAMC2) is highly expressed in anaplastic thyroid carcinoma and is...

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