Let-7a/cMyc/CCAT1/miR-17-5p Circuit Re-sensitizes Atezolizumab Resistance in Triple Negative Breast Cancer through Modulating PD-L1.

Abstract

BACKGROUND:

Triple negative breast cancer (TNBC) is an immunogenically hot tumor. The immune checkpoint blockades (ICBs) have been recently emerged as promising therapeutic candidates for several malignancies including TNBC. Yet, the development of innate and/or adaptive resistance by TNBC patients towards ICBs such as programmed death-ligand 1 (PD-L1) inhibitors (e.g. Atezolizumab) shed the light on importance of identifying the underlying mechanisms regulating PD-L1 in TNBC. Recently, it was reported that non-coding RNAs (ncRNAs) perform a fundamental role in regulating PD-L1 expression in TNBC. Hence, this study aims to explore a novel ncRNA axis tuning PD-L1 in TNBC patients and investigate its possible involvement in fighting Atezolizumab resistance.

METHODS:

In-silico screening was executed to identify ncRNAs that could potentially target PD-L1. Screening of PD-L1 and the nominated ncRNAs (miR-17-5p, let-7a and CCAT1 lncRNA) was performed in BC patients and cell lines. Ectopic expression and/or knockdown of respective ncRNAs were performed in MDA-MB-231. Cellular viability, migration and clonogenic capacities were evaluated using MTT, scratch assay and colony-forming assay, respectively.

RESULTS:

PD-L1 was upregulated in BC patients, especially in TNBC patients. PD-L1 is positively associated with lymph node metastasis and high Ki-67 in recruited BC patients. Let-7a and miR-17-5p were nominated as potential regulators of PD-L1. Ectopic expression of let-7a and miR-17-5p caused a noticeable reduction in PD-L1 levels in TNBC cells. In order to investigate the whole ceRNA circuit regulating PD-L1 in TNBC, intensive bioinformatic studies were performed. The lncRNA, Colon Cancer-associated transcript 1 (CCAT1), was reported to target PD-L1 regulating miRNAs. Results showed that CCAT1 is an upregulated oncogenic lncRNA in TNBC patients and cell lines. CCAT1 siRNAs induced a noticeable reduction in PD-L1 levels and a marked increase in miR-17-5p level, building up a novel regulatory axis CCAT1/miR-17-5p/PD-L1 in TNBC cells that was tuned by the let-7a/c-Myc engine. On the functional level, co-treatment of CCAT-1 siRNAs and let-7a mimics efficiently relieved Atezolizumab resistance in MDA-MB-231 cells.

CONCLUSION:

The present study revealed a novel PD-L1 regulatory axis via targeting let-7a/c-Myc/CCAT/miR-17-5p. Additionally, it sheds the light on the potential combinational role of CCAT-1 siRNAs and Let-7a mimics in relieving Atezolizumab resistance in TNBC patients.

• Authors: Noha A Selem, Heba Nafae, Tamer Manie, Rana A Youness, Mohamed Z Gad
• Journal: Pathology, research and practice (Pathol Res Pract) Vol. 248 Pg. 154579 (Aug 2023) ISSN: 1618-0631 [Electronic] Germany
• PMID: 37301086 (Publication Type: Journal Article)
• Copyright: Copyright © 2023 Elsevier GmbH. All rights reserved.

Chemical References

• atezolizumab
• B7-H1 Antigen
• CCAT1 long noncoding RNA, human
• CD274 protein, human
• MicroRNAs
• MIRN17 microRNA, human
• RNA, Long Noncoding

Topics

• Humans
• B7-H1 Antigen (genetics, metabolism)
• Cell Line, Tumor
• Cell Proliferation (genetics)
• Colonic Neoplasms (genetics)
• Gene Expression Regulation, Neoplastic (genetics)
• MicroRNAs (metabolism)
• RNA, Long Noncoding (genetics)
• Triple Negative Breast Neoplasms (drug therapy, genetics, metabolism)