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The molecular basis of 7SK RNA methylation in non-small cell lung cancer

Award Number: R21CA287107
ORGANIZATION: NATIONAL CANCER INSTITUTE
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 01/01/2024
PERIOD OF PERFORMANCE END DATE: 12/31/2025

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The molecular basis of 7SK RNA methylation in non-small cell lung cancer - PROJECT SUMMARY Lung cancer is a leading cause of cancer death in the United States, with non-small cell lung cancer (NSCLC) being the major subtype accounting for approximately 85% of cases. Effective treatments for NSCLC are currently lacking due to the complexity of its molecular subtypes and the incomplete understanding of its underlying mechanisms. RNA N6-methyladenosine (m6A) modification is increasingly recognized as being deregulated in human cancers, however, specific m6A events in NSCLC are yet to be identified. We found elevated m6A modification on 7SK small nuclear RNA in NSCLC cells and a NSCLC mouse model. Targeted reduction of m6A-7SK inhibits NSCLC cell growth in soft agar colony formation assays. 7SK regulates RNA Polymerase II (Pol II) transcription and is a central player in controlling Pol II transcriptional pausing and elongation. Therefore, m6A modification of 7SK may underlie NSCLC formation and phenotype. The goal of this study is to dissect the molecular pathways that regulate m6A-7SK and decipher the mechanisms by which m6A-7SK affects gene regulatory networks in NSCLC. The first aim is to identify the m6A sites using real-time PCR-based m6A detection assay, and to identify writer and eraser enzymes of 7SK by short hairpin RNA knockdown. The second aim is to determine how m6A-7SK influences gene expression and NSCLC formation. CRISPR dCasRx-ALKBH5 fusion protein will be used to modulate m6A-7SK in NSCLC cells, and cell proliferation and migration assays will be employed. To determine the molecular details of m6A-7SK-mediated gene regulation, SHAPE-MaP sequencing will probe the structure of m6A-7SK; nascent RNA-seq, ChIP-seq of Pol II, and ChIRP-seq of 7SK will be performed to survey transcriptional program in NSCLC cells with different levels of m6A-7SK. The results of this study will reveal novel molecular details of m6A modifications on 7SK and how they regulate cancer gene expression. This information will provide the basis for future research to develop novel NSCLC therapeutics by altering RNA m6A levels.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2025 ( Subtotal = $291,209 )
2025	2025	UNIVERSITY OF FLORIDA	1523 UNION RD RM 207	GAINESVILLE	FL	32611	ALACHUA	USA	Cancer Biology Research	000	2	12/9/2024	NON-COMPETING CONTINUATION	$192,493
2025	2025	UNIVERSITY OF FLORIDA	1523 UNION RD RM 207	GAINESVILLE	FL	32611	ALACHUA	USA	Cancer Biology Research	001	2	12/16/2024	SUPPLEMENT FOR EXPANSION	$77,328
2025	2025	UNIVERSITY OF FLORIDA	1523 UNION RD RM 207	GAINESVILLE	FL	32611	ALACHUA	USA	Cancer Biology Research	002	2	6/12/2025	NON-COMPETING CONTINUATION	$21,388
														Subtotal = $291,209

Issue Date FY: 2024 ( Subtotal = $230,861 )
2024	2024	UNIVERSITY OF FLORIDA	1523 UNION RD RM 207	GAINESVILLE	FL	32611	ALACHUA	USA	Cancer Biology Research	001	1	8/5/2024	SUPPLEMENT FOR EXPANSION	$52,627
2024	2024	UNIVERSITY OF FLORIDA	1523 UNION RD RM 207	GAINESVILLE	FL	32611	ALACHUA	USA	Cancer Biology Research	000	1	12/14/2023	NEW	$178,234
														Subtotal = $230,861

Grand Total All Awards = $522,070

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The molecular basis of 7SK RNA methylation in non-small cell lung cancer

Award Number: R21CA287107

ORGANIZATION: NATIONAL CANCER INSTITUTE

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 01/01/2024

PERIOD OF PERFORMANCE END DATE: 12/31/2025

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