Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer
Introduction: Breast cancer is the most common cancer in women, with roughly 10–15% of new cases classified as triple-negative breast cancer (TNBC). Traditional chemotherapies are often toxic to normal cells. Therefore, it is important to discover new anticancer compounds that target TNBC while caus...
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| Format: | Article |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1243763/full |
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| author | Shradheya R. R. Gupta Tram M. Ta Maryam Khan Archana Singh Indrakant K. Singh Indrakant K. Singh Indrakant K. Singh Bela Peethambaran |
| author_facet | Shradheya R. R. Gupta Tram M. Ta Maryam Khan Archana Singh Indrakant K. Singh Indrakant K. Singh Indrakant K. Singh Bela Peethambaran |
| author_sort | Shradheya R. R. Gupta |
| collection | DOAJ |
| description | Introduction: Breast cancer is the most common cancer in women, with roughly 10–15% of new cases classified as triple-negative breast cancer (TNBC). Traditional chemotherapies are often toxic to normal cells. Therefore, it is important to discover new anticancer compounds that target TNBC while causing minimal damage to normal cells. Receptor tyrosine kinase-like Orphan Receptor 1 (ROR1) is an oncofetal protein overexpressed in numerous human malignancies, including TNBC. This study investigated potential small molecules targeting ROR1.Methodology: Using AutoDock Vina and Glide, we screened 70,000 chemicals for our investigation. We obtained 10 representative compounds via consensus voting, deleting structural alerts, and clustering. After manual assessment, compounds 2 and 4 were chosen for MD simulation and cell viability experiment. Compound 4 showed promising results in the viability assay, which led us to move further with the apoptosis assay and immunoblotting.Results: Compound 4 (CID1261330) had docking scores of −6.635 and −10.8. It fits into the pocket and shows interactions with GLU64, ASP174, and PHE93. Its RMSD fluctuates around 0.20 nm and forms two stable H-bonds indicating compound 4 stability. It inhibits cell proliferation in MDA-MB-231, HCC1937, and HCC1395 cell lines, with IC50 values of approximately 2 μM to 10 μM, respectively. Compound 4 did not kill non-malignant epithelial breast cells MCF-10A (IC50 > 27 μM). These results were confirmed by the significant number of apoptotic cells in MDA-MB-231 cells (47.6%) but not in MCF-10A cells (7.3%). Immunoblot analysis provided additional support in the same direction.Discussion: These findings collectively suggest that compound 4 has the potential to effectively eliminate TNBC cells while causing minimal harm to normal breast cells. The promising outcomes of this study lay the groundwork for further testing of compound 4 in other malignancies characterized by ROR1 upregulation, serving as a proof-of-concept for its broader applicability. |
| first_indexed | 2024-03-12T01:13:50Z |
| format | Article |
| id | doaj.art-81a0d7c3f84c410da28b4f272dfe0101 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-12T01:13:50Z |
| publishDate | 2023-09-01 |
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| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-81a0d7c3f84c410da28b4f272dfe01012023-09-13T21:49:53ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-09-011110.3389/fcell.2023.12437631243763Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancerShradheya R. R. Gupta0Tram M. Ta1Maryam Khan2Archana Singh3Indrakant K. Singh4Indrakant K. Singh5Indrakant K. Singh6Bela Peethambaran7Molecular Biology Research Laboratory, Department of Zoology, Deshbandhu College, University of Delhi, New Delhi, IndiaDepartment of Biology, Saint Joseph’s University, Philadelphia, PA, United StatesDepartment of Biology, Saint Joseph’s University, Philadelphia, PA, United StatesDepartment of Botany, Hans Raj College, University of Delhi, New Delhi, IndiaMolecular Biology Research Laboratory, Department of Zoology, Deshbandhu College, University of Delhi, New Delhi, IndiaDelhi School of Public Health, Institute of Eminence, University of Delhi, New Delhi, IndiaNorris Comprehensive Cancer Center, Division of Medical Oncology, University of Southern California, Los Angeles, CA, United StatesDepartment of Biology, Saint Joseph’s University, Philadelphia, PA, United StatesIntroduction: Breast cancer is the most common cancer in women, with roughly 10–15% of new cases classified as triple-negative breast cancer (TNBC). Traditional chemotherapies are often toxic to normal cells. Therefore, it is important to discover new anticancer compounds that target TNBC while causing minimal damage to normal cells. Receptor tyrosine kinase-like Orphan Receptor 1 (ROR1) is an oncofetal protein overexpressed in numerous human malignancies, including TNBC. This study investigated potential small molecules targeting ROR1.Methodology: Using AutoDock Vina and Glide, we screened 70,000 chemicals for our investigation. We obtained 10 representative compounds via consensus voting, deleting structural alerts, and clustering. After manual assessment, compounds 2 and 4 were chosen for MD simulation and cell viability experiment. Compound 4 showed promising results in the viability assay, which led us to move further with the apoptosis assay and immunoblotting.Results: Compound 4 (CID1261330) had docking scores of −6.635 and −10.8. It fits into the pocket and shows interactions with GLU64, ASP174, and PHE93. Its RMSD fluctuates around 0.20 nm and forms two stable H-bonds indicating compound 4 stability. It inhibits cell proliferation in MDA-MB-231, HCC1937, and HCC1395 cell lines, with IC50 values of approximately 2 μM to 10 μM, respectively. Compound 4 did not kill non-malignant epithelial breast cells MCF-10A (IC50 > 27 μM). These results were confirmed by the significant number of apoptotic cells in MDA-MB-231 cells (47.6%) but not in MCF-10A cells (7.3%). Immunoblot analysis provided additional support in the same direction.Discussion: These findings collectively suggest that compound 4 has the potential to effectively eliminate TNBC cells while causing minimal harm to normal breast cells. The promising outcomes of this study lay the groundwork for further testing of compound 4 in other malignancies characterized by ROR1 upregulation, serving as a proof-of-concept for its broader applicability.https://www.frontiersin.org/articles/10.3389/fcell.2023.1243763/fullbreast cancerTNBCRor1kinaseMCF-10AMDA-MB-231 |
| spellingShingle | Shradheya R. R. Gupta Tram M. Ta Maryam Khan Archana Singh Indrakant K. Singh Indrakant K. Singh Indrakant K. Singh Bela Peethambaran Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer Frontiers in Cell and Developmental Biology breast cancer TNBC Ror1 kinase MCF-10A MDA-MB-231 |
| title | Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer |
| title_full | Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer |
| title_fullStr | Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer |
| title_full_unstemmed | Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer |
| title_short | Identification and validation of a small molecule targeting ROR1 for the treatment of triple negative breast cancer |
| title_sort | identification and validation of a small molecule targeting ror1 for the treatment of triple negative breast cancer |
| topic | breast cancer TNBC Ror1 kinase MCF-10A MDA-MB-231 |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1243763/full |
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