Proceedings

Validating hit genes from CRISPR screening on response toward combinations of targeted therapeutic drugs in acute myeloid leukemia (AML)

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Acute myeloid leukemia (AML) is a heterogenous blood cancer disease that develops from the stem cell precursors which the myeloid lineage happened due to genetic lesion that leads to overproduction and failure to differentiate these myeloid blast cells. A mutation in the FMS-like tyrosine kinase 3 (FLT3) gene, is the most frequent genetic lesion in AML patients, this mutation is linked with a poor prognosis. These make FLT3 a small molecular target for AML treatment. Gilteritinib as an FLT3 inhibitor drug may increase the survival of AML with FLT3 mutations. Moreover, the benefit is short-term due to the development of resistance. Therefore, to improve AML treatment FLT3 inhibitors were combined with a BCL2 inhibitor, venetoclax, and a MEK1/2 inhibitor, trametinib. In a preliminary study, a whole genome CRISPR knock-out (KO) screen was used to check whether these combination drug strategies will benefit every AML subtype and not cause resistance. Through the CRISPR KO screen, AHR/ARNT genes were identified to be involved in the response to the combination of gilteritinib and venetoclax, and the PTPN1 gene was involved in the response to the combination of gilteritinib and trametinib. In this study, we validated whether those hit genes cause resistance to the gilteritinib-based combination strategies in the MOLM13 AML cell line, and use molecular cloning techniques to generate hit genes KO cell. Then, examine the efficiency of knock out cell through western blot and sequencing techniques, and validate drug responses using MTT assay. From our result, it shows that AHR/ARNT KO in response to the combination of gilteritinib and venetoclax is inconclusive due to several problem encounters, while PTPN1 KO is shown resistance to the combination of gilteritinib and trametinib which affect the treatment option for AML patient in the future.

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Detail Information

Series Title

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Call Number

BM 23-031

Publisher

i3L, Jakarta : i3L, Jakarta., 2023

Collation

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Language

English

ISBN/ISSN

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Classification

NONE

Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

Acute Myeloid Leukemia
Drug Combination
CRISPR Screening
Drug Resistance
Targeted therapy

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