Summary

Myelodysplastic syndromes (MDS) are a heterogeneous hematopoietic neoplasm driven by somatically acquired genetic mutations and epigenetic alterations. Traditionally, diagnosis of MDS is based on the presence of dysplasia in one or more hematopoietic lineages and /or the presence of characteristic chromosomal abnormalities. Since some of dysplastic changes are common to other myeloid malignancies and majority of patients with MDS present with a normal karyotype, diagnosis of MDS by routine diagnostic approaches has been complicated. Therefore, detection of these anomalies needs several molecular techniques such as conventional cytogenetics, FISH, ARMS PCR somatic mutation analysis assays, comparative genomic hybridization, copy number variation analysis, sequencing etc., with different resolutions and sensitivities. However, the use of these molecular techniques should be in line with a rational approach initially by focusing on structural and numerical chromosomal anomalies, subsequently mutations of vital genes in affected chromosomes. It has been observed that the existence of these molecular anomalies is not consistent, unique, and evolves continuously, thus making interpretation of MDS tests more challenging. The present study focuses on retrieving a higher number of analyzable cells in a bone marrow aspiration sample transported using an in-house developed bone marrow transport medium, thus facilitating an extensive molecular analysis by conventional karyotyping, FISH and somatic mutation analysis of selected vital genes associated with risk stratification.

Objectives

General Objective:

Evaluate the outcomes of molecular analysis of bone marrow aspirates obtained from clinically diagnosed patients with MDS in their prognostic risk stratification using an in-house developed bone marrow transporting medium.

Specific Objectives:

1. A comprehensive study on the stability and sterility of in-house developed bone marrow transporting

medium

2. Study the effect of in-house developed bone marrow transporting medium on the direct harvest of

unstimulated bone marrow aspirates

3. Compare the outcomes of aspirates of bone marrow culture transported in, in-house developed bone

marrow transport medium vs commercially available sodium heparin containing vacutainers.

4. Compare the outcomes of conventional karyotypes of unstimulated bone marrow cultures with that

of the outcome of their meta-phase FISH analysis targeting Del(5q), Del (7q), Del(20qter), and

Trisomy 8.

5. Validation of MDS FISH panels; Del(5q), Del (7q), Del(20qter), and Trisomy 8
6. Mutation analysis of two vital genes associated with risk stratification of MDS; TP53, and TET2

Major Equipment Facilitated by Grant