Faculty Appointments
Assistant Professor of Medicine
Assistant Professor of Cell & Developmental Biology
Education
Ph.D., Molecular/Cancer Biology, Max Planck Institute of Biochemistry, Munich, GermanyM.Sc., Biomedical Sciences, University of Delhi, Delhi, IndiaB.Sc., University of Delhi, Delhi, India
Office Address
2213 Garland Avenue
10415A, MRB-IV
Nashville, TN 37232-0441
10415A, MRB-IV
Nashville, TN 37232-0441
Research Description
Our lab studies receptor tyrosine kinase (RTK) signaling in human health and disease with an emphasis on epidermal growth factor receptor (EGFR) signaling. The research may broadly be divided into two areas:
1. Trafficking of EGFR ligands in polarized epithelial cells: Spatial cues to regulate EGFR signaling are best appreciated in three-dimensional (3D) cultures (e.g. Transwell, type I collagen, and Matrigel cultures) than conventional 2D plastic cultures, where EGFR ligands are delivered and secreted in a polarized manner (apical vs basolateral trafficking). A major focus within this area is to understand the loss of epithelial polarity induced by loss of polarized trafficking of the EGFR ligand, epiregulin, and role of novel epiregulin-interacting proteins in the observed phenotypes.
2. Cetuximab resistance in colorectal cancer: Using 3D cultures of colorectal cancer (CRC) cells, we study novel resistance mechanisms to cetuximab, which is an FDA-approved EGFR neutralizing monoclonal antibody. We have discovered two non-genetic modes of cetuximab resistance and identified means to overcome this resistance by addition of a dual MET/RON RTK inhibitor, crizotinib. We are committed to elucidating the mechanistic underpinnings of this mode of cetuximab resistance, its scope in human CRC, and the therapeutic benefit of combined blockade of EGFR with RTKs, MET and RON.
This multi-disciplinary and collaborative research integrates approaches like molecular cell biology, live or fixed (confocal) microscopy, comparative RNA-seq (single-cell or bulk), proteomic analysis, pre-clinical models (patient-derived xenografts and organoids, PDXs/PDOs), and small-animal imaging.
The research program is supported by the NIH/NCI CA248505-01A1 and American Cancer Society RSG-20-130-01-DDC grants.
1. Trafficking of EGFR ligands in polarized epithelial cells: Spatial cues to regulate EGFR signaling are best appreciated in three-dimensional (3D) cultures (e.g. Transwell, type I collagen, and Matrigel cultures) than conventional 2D plastic cultures, where EGFR ligands are delivered and secreted in a polarized manner (apical vs basolateral trafficking). A major focus within this area is to understand the loss of epithelial polarity induced by loss of polarized trafficking of the EGFR ligand, epiregulin, and role of novel epiregulin-interacting proteins in the observed phenotypes.
2. Cetuximab resistance in colorectal cancer: Using 3D cultures of colorectal cancer (CRC) cells, we study novel resistance mechanisms to cetuximab, which is an FDA-approved EGFR neutralizing monoclonal antibody. We have discovered two non-genetic modes of cetuximab resistance and identified means to overcome this resistance by addition of a dual MET/RON RTK inhibitor, crizotinib. We are committed to elucidating the mechanistic underpinnings of this mode of cetuximab resistance, its scope in human CRC, and the therapeutic benefit of combined blockade of EGFR with RTKs, MET and RON.
This multi-disciplinary and collaborative research integrates approaches like molecular cell biology, live or fixed (confocal) microscopy, comparative RNA-seq (single-cell or bulk), proteomic analysis, pre-clinical models (patient-derived xenografts and organoids, PDXs/PDOs), and small-animal imaging.
The research program is supported by the NIH/NCI CA248505-01A1 and American Cancer Society RSG-20-130-01-DDC grants.
Research Keywords
Colorectal cancer; epithelial polarity; transformation; receptor tyrosine kinases (RTKs) and cognate ligands with special emphasis on ERBBs, MET, and RON; protein trafficking; resistance to EGFR-targeted therapies; Cancer; Cetuximab
Publications
Snyder RA, Burtness B, Cho M, Del Rivero J, Doroshow DB, Hitchcock KE, Kalyan A, Kim CA, Lukovic J, Parikh AR, Sanford NN, Singh B, Shen C, Shroff RT, Vijayvergia N, Goodman KA, Kunz PL. The Room Where It Happens: Addressing Diversity, Equity, and Inclusion in NCTN Clinical Trial Leadership. J Natl Cancer Inst [print-electronic]. 2023 Jun 6/26/2023; PMID: 37364007, PII: 7207830, DOI: 10.1093/jnci/djad121, ISSN: 1460-2105.
Cao T, Lu Y, Wang Q, Qin H, Li H, Guo H, Ge M, Glass SE, Singh B, Zhang W, Dong J, Du F, Qian A, Tian Y, Wang X, Li C, Wu K, Fan D, Nie Y, Coffey RJ, Zhao X. A CGA/EGFR/GATA2 positive feedback circuit confers chemoresistance in gastric cancer. J Clin Invest. 2022 Mar 3/15/2022; 132(6): PMID: 35289315, PMCID: PMC8920335, PII: 154074, DOI: 10.1172/JCI154074, ISSN: 1558-8238.
Singh B, Bogatcheva G, Krystofiak E, McKinley ET, Hill S, Rose KL, Higginbotham JN, Coffey RJ. Induction of apically mistrafficked epiregulin disrupts epithelial polarity via aberrant EGFR signaling. J Cell Sci [print-electronic]. 2021 Sep 9/15/2021; 134(18): PMID: 34406412, PMCID: PMC8466005, PII: 271860, DOI: 10.1242/jcs.255927, ISSN: 1477-9137.
Starchenko A, Graves-Deal R, Brubaker D, Li C, Yang Y, Singh B, Coffey RJ, Lauffenburger DA. Cell surface integrin a5ß1 clustering negatively regulates receptor tyrosine kinase signaling in colorectal cancer cells via glycogen synthase kinase 3. Integr Biol (Camb). 2021 Jun 6/15/2021; 13(6): 153-66. PMID: 34037774, PMCID: PMC8204629, PII: 6284376, DOI: 10.1093/intbio/zyab009, ISSN: 1757-9708.
Wen X, Ou YC, Bogatcheva G, Thomas G, Mahadevan-Jansen A, Singh B, Lin EC, Bardhan R. Probing metabolic alterations in breast cancer in response to molecular inhibitors with Raman spectroscopy and validated with mass spectrometry. Chem Sci. 2020 Aug 8/20/2020; 11(36): 9863-74. PMID: 34094246, PMCID: PMC8162119, PII: d0sc02221g, DOI: 10.1039/d0sc02221g, ISSN: 2041-6520.
Cao Z, Singh B, Li C, Markham NO, Carrington LJ, Franklin JL, Graves-Deal R, Kennedy EJ, Goldenring JR, Coffey RJ. Protein kinase A-mediated phosphorylation of naked cuticle homolog 2 stimulates cell-surface delivery of transforming growth factor-a for epidermal growth factor receptor transactivation. Traffic. 2019 May; 20(5): 357-68. PMID: 30941853, PMCID: PMC6618044, DOI: 10.1111/tra.12642, ISSN: 1600-0854.
Graves-Deal R, Bogatcheva G, Rehman S, Lu Y, Higginbotham JN, Singh B. Broad-spectrum receptor tyrosine kinase inhibitors overcome de novo and acquired modes of resistance to EGFR-targeted therapies in colorectal cancer. Oncotarget. 2019 Feb 2/12/2019; 10(13): 1320-33. PMID: 30863492, PMCID: PMC6407678, PII: 26663, DOI: 10.18632/oncotarget.26663, ISSN: 1949-2553.
Lu Y, Zhao X, Liu Q, Li C, Graves-Deal R, Cao Z, Singh B, Franklin JL, Wang J, Hu H, Wei T, Yang M, Yeatman TJ, Lee E, Saito-Diaz K, Hinger S, Patton JG, Chung CH, Emmrich S, Klusmann JH, Fan D, Coffey RJ. LncRNA MIR100HG-derived miR-100 and miR-125b mediate cetuximab resistance via Wnt/ß-catenin signaling. Nat. Med [print-electronic]. 2017 Nov; 23(11): 1331-41. PMID: 29035371, PMCID: PMC5961502, PII: nm.4424, DOI: 10.1038/nm.4424, ISSN: 1546-170X.
Li C, Singh B, Graves-Deal R, Ma H, Starchenko A, Fry WH, Lu Y, Wang Y, Bogatcheva G, Khan MP, Milne GL, Zhao S, Ayers GD, Li N, Hu H, Washington MK, Yeatman TJ, McDonald OG, Liu Q, Coffey RJ. Three-dimensional culture system identifies a new mode of cetuximab resistance and disease-relevant genes in colorectal cancer. Proc. Natl. Acad. Sci. U.S.A [print-electronic]. 2017 Apr 4/4/2017; 114(14): E2852-E2861. PMID: 28320945, PMCID: PMC5389279, PII: 1618297114, DOI: 10.1073/pnas.1618297114, ISSN: 1091-6490.
Starchenko A, Graves-Deal R, Yang YP, Li C, Zent R, Singh B, Coffey RJ. Clustering of Integrin alpha 5 at the lateral membrane restores epithelial polarity in invasive colorectal cancer cells. Mol. Biol. Cell [print-electronic]. 2017 Mar 3/29/2017; PMID: 28356422, PII: mbc.E16-12-0852, DOI: 10.1091/mbc.E16-12-0852, ISSN: 1939-4586.
Singh B, Bogatcheva G, Starchenko A, Sinnaeve J, Lapierre LA, Williams JA, Goldenring JR, Coffey RJ. Induction of lateral lumens through disruption of a monoleucine-based basolateral-sorting motif in betacellulin. J. Cell. Sci [print-electronic]. 2015 Sep 9/15/2015; 128(18): 3444-55. PMID: 26272915, PMCID: PMC4582402, PII: jcs.170852, DOI: 10.1242/jcs.170852, ISSN: 1477-9137.
Singh B, Coffey RJ. From wavy hair to naked proteins: the role of transforming growth factor alpha in health and disease. Semin. Cell Dev. Biol [print-electronic]. 2014 Apr; 28: 12-21. PMID: 24631356, PMCID: PMC4105142, PII: S1084-9521(14)00031-7, DOI: 10.1016/j.semcdb.2014.03.003, ISSN: 1096-3634.
Singh B, Coffey RJ. Trafficking of epidermal growth factor receptor ligands in polarized epithelial cells. Annu. Rev. Physiol [print-electronic]. 2014; 76: 275-300. PMID: 24215440, PMCID: PMC4180094, DOI: 10.1146/annurev-physiol-021113-170406, ISSN: 1545-1585.
Singh B, Bogatcheva G, Washington MK, Coffey RJ. Transformation of polarized epithelial cells by apical mistrafficking of epiregulin. Proc. Natl. Acad. Sci. U.S.A [print-electronic]. 2013 May 5/28/2013; 110(22): 8960-5. PMID: 23671122, PMCID: PMC3670353, PII: 1305508110, DOI: 10.1073/pnas.1305508110, ISSN: 1091-6490.
Gephart JD, Singh B, Higginbotham JN, Franklin JL, Gonzalez A, Fölsch H, Coffey RJ. Identification of a novel mono-leucine basolateral sorting motif within the cytoplasmic domain of amphiregulin. Traffic [print-electronic]. 2011 Dec; 12(12): 1793-804. PMID: 21917092, PMCID: PMC3886124, DOI: 10.1111/j.1600-0854.2011.01282.x, ISSN: 1600-0854.
Singh B, Schneider M, Knyazev P, Ullrich A. UV-induced EGFR signal transactivation is dependent on proligand shedding by activated metalloproteases in skin cancer cell lines. Int. J. Cancer. 2009 Feb 2/1/2009; 124(3): 531-9. PMID: 19003995, DOI: 10.1002/ijc.23974, ISSN: 1097-0215.
Chopra P, Singh B, Singh R, Vohra R, Koul A, Meena LS, Koduri H, Ghildiyal M, Deol P, Das TK, Tyagi AK, Singh Y. Phosphoprotein phosphatase of Mycobacterium tuberculosis dephosphorylates serine-threonine kinases PknA and PknB. Biochem. Biophys. Res. Commun. 2003 Nov 11/7/2003; 311(1): 112-20. PMID: 14575702, PII: S0006291X03020011, ISSN: 0006-291X.
Cao T, Lu Y, Wang Q, Qin H, Li H, Guo H, Ge M, Glass SE, Singh B, Zhang W, Dong J, Du F, Qian A, Tian Y, Wang X, Li C, Wu K, Fan D, Nie Y, Coffey RJ, Zhao X. A CGA/EGFR/GATA2 positive feedback circuit confers chemoresistance in gastric cancer. J Clin Invest. 2022 Mar 3/15/2022; 132(6): PMID: 35289315, PMCID: PMC8920335, PII: 154074, DOI: 10.1172/JCI154074, ISSN: 1558-8238.
Singh B, Bogatcheva G, Krystofiak E, McKinley ET, Hill S, Rose KL, Higginbotham JN, Coffey RJ. Induction of apically mistrafficked epiregulin disrupts epithelial polarity via aberrant EGFR signaling. J Cell Sci [print-electronic]. 2021 Sep 9/15/2021; 134(18): PMID: 34406412, PMCID: PMC8466005, PII: 271860, DOI: 10.1242/jcs.255927, ISSN: 1477-9137.
Starchenko A, Graves-Deal R, Brubaker D, Li C, Yang Y, Singh B, Coffey RJ, Lauffenburger DA. Cell surface integrin a5ß1 clustering negatively regulates receptor tyrosine kinase signaling in colorectal cancer cells via glycogen synthase kinase 3. Integr Biol (Camb). 2021 Jun 6/15/2021; 13(6): 153-66. PMID: 34037774, PMCID: PMC8204629, PII: 6284376, DOI: 10.1093/intbio/zyab009, ISSN: 1757-9708.
Wen X, Ou YC, Bogatcheva G, Thomas G, Mahadevan-Jansen A, Singh B, Lin EC, Bardhan R. Probing metabolic alterations in breast cancer in response to molecular inhibitors with Raman spectroscopy and validated with mass spectrometry. Chem Sci. 2020 Aug 8/20/2020; 11(36): 9863-74. PMID: 34094246, PMCID: PMC8162119, PII: d0sc02221g, DOI: 10.1039/d0sc02221g, ISSN: 2041-6520.
Cao Z, Singh B, Li C, Markham NO, Carrington LJ, Franklin JL, Graves-Deal R, Kennedy EJ, Goldenring JR, Coffey RJ. Protein kinase A-mediated phosphorylation of naked cuticle homolog 2 stimulates cell-surface delivery of transforming growth factor-a for epidermal growth factor receptor transactivation. Traffic. 2019 May; 20(5): 357-68. PMID: 30941853, PMCID: PMC6618044, DOI: 10.1111/tra.12642, ISSN: 1600-0854.
Graves-Deal R, Bogatcheva G, Rehman S, Lu Y, Higginbotham JN, Singh B. Broad-spectrum receptor tyrosine kinase inhibitors overcome de novo and acquired modes of resistance to EGFR-targeted therapies in colorectal cancer. Oncotarget. 2019 Feb 2/12/2019; 10(13): 1320-33. PMID: 30863492, PMCID: PMC6407678, PII: 26663, DOI: 10.18632/oncotarget.26663, ISSN: 1949-2553.
Lu Y, Zhao X, Liu Q, Li C, Graves-Deal R, Cao Z, Singh B, Franklin JL, Wang J, Hu H, Wei T, Yang M, Yeatman TJ, Lee E, Saito-Diaz K, Hinger S, Patton JG, Chung CH, Emmrich S, Klusmann JH, Fan D, Coffey RJ. LncRNA MIR100HG-derived miR-100 and miR-125b mediate cetuximab resistance via Wnt/ß-catenin signaling. Nat. Med [print-electronic]. 2017 Nov; 23(11): 1331-41. PMID: 29035371, PMCID: PMC5961502, PII: nm.4424, DOI: 10.1038/nm.4424, ISSN: 1546-170X.
Li C, Singh B, Graves-Deal R, Ma H, Starchenko A, Fry WH, Lu Y, Wang Y, Bogatcheva G, Khan MP, Milne GL, Zhao S, Ayers GD, Li N, Hu H, Washington MK, Yeatman TJ, McDonald OG, Liu Q, Coffey RJ. Three-dimensional culture system identifies a new mode of cetuximab resistance and disease-relevant genes in colorectal cancer. Proc. Natl. Acad. Sci. U.S.A [print-electronic]. 2017 Apr 4/4/2017; 114(14): E2852-E2861. PMID: 28320945, PMCID: PMC5389279, PII: 1618297114, DOI: 10.1073/pnas.1618297114, ISSN: 1091-6490.
Starchenko A, Graves-Deal R, Yang YP, Li C, Zent R, Singh B, Coffey RJ. Clustering of Integrin alpha 5 at the lateral membrane restores epithelial polarity in invasive colorectal cancer cells. Mol. Biol. Cell [print-electronic]. 2017 Mar 3/29/2017; PMID: 28356422, PII: mbc.E16-12-0852, DOI: 10.1091/mbc.E16-12-0852, ISSN: 1939-4586.
Singh B, Bogatcheva G, Starchenko A, Sinnaeve J, Lapierre LA, Williams JA, Goldenring JR, Coffey RJ. Induction of lateral lumens through disruption of a monoleucine-based basolateral-sorting motif in betacellulin. J. Cell. Sci [print-electronic]. 2015 Sep 9/15/2015; 128(18): 3444-55. PMID: 26272915, PMCID: PMC4582402, PII: jcs.170852, DOI: 10.1242/jcs.170852, ISSN: 1477-9137.
Singh B, Coffey RJ. From wavy hair to naked proteins: the role of transforming growth factor alpha in health and disease. Semin. Cell Dev. Biol [print-electronic]. 2014 Apr; 28: 12-21. PMID: 24631356, PMCID: PMC4105142, PII: S1084-9521(14)00031-7, DOI: 10.1016/j.semcdb.2014.03.003, ISSN: 1096-3634.
Singh B, Coffey RJ. Trafficking of epidermal growth factor receptor ligands in polarized epithelial cells. Annu. Rev. Physiol [print-electronic]. 2014; 76: 275-300. PMID: 24215440, PMCID: PMC4180094, DOI: 10.1146/annurev-physiol-021113-170406, ISSN: 1545-1585.
Singh B, Bogatcheva G, Washington MK, Coffey RJ. Transformation of polarized epithelial cells by apical mistrafficking of epiregulin. Proc. Natl. Acad. Sci. U.S.A [print-electronic]. 2013 May 5/28/2013; 110(22): 8960-5. PMID: 23671122, PMCID: PMC3670353, PII: 1305508110, DOI: 10.1073/pnas.1305508110, ISSN: 1091-6490.
Gephart JD, Singh B, Higginbotham JN, Franklin JL, Gonzalez A, Fölsch H, Coffey RJ. Identification of a novel mono-leucine basolateral sorting motif within the cytoplasmic domain of amphiregulin. Traffic [print-electronic]. 2011 Dec; 12(12): 1793-804. PMID: 21917092, PMCID: PMC3886124, DOI: 10.1111/j.1600-0854.2011.01282.x, ISSN: 1600-0854.
Singh B, Schneider M, Knyazev P, Ullrich A. UV-induced EGFR signal transactivation is dependent on proligand shedding by activated metalloproteases in skin cancer cell lines. Int. J. Cancer. 2009 Feb 2/1/2009; 124(3): 531-9. PMID: 19003995, DOI: 10.1002/ijc.23974, ISSN: 1097-0215.
Chopra P, Singh B, Singh R, Vohra R, Koul A, Meena LS, Koduri H, Ghildiyal M, Deol P, Das TK, Tyagi AK, Singh Y. Phosphoprotein phosphatase of Mycobacterium tuberculosis dephosphorylates serine-threonine kinases PknA and PknB. Biochem. Biophys. Res. Commun. 2003 Nov 11/7/2003; 311(1): 112-20. PMID: 14575702, PII: S0006291X03020011, ISSN: 0006-291X.
Available Postdoctoral Position Details
Posted: 7/2/2021
We are looking for postdoctoral candidates to participate in basic studies controlling epithelial polarity as well as more translational efforts to develop innovative therapeutic approaches for colorectal cancer.
Available projects include:
- Regulation of EGFR signaling by polarized trafficking of its ligand epiregulin and mechanisms underlying transformation upon apical mistrafficking of epiregulin;
- Role of receptor tyrosine kinases MET and RON in resistance to EGFR-targeted therapeutics in colorectal cancer.
Please send CV and a cover letter to bhuminder.singh@vumc.org
Thank you
Bhumi