HER2 reactivation through acquisition of the HER2 L755S mutation as a mechanism of acquired resistance to HER2-targeted therapy in HER2+ breast cancer

Xiaowei Xu, Carmine De Angelis, Kathleen A. Burke, Agostina Nardone, Huizhong Hu, Lanfang Qin, Jamunarani Veeraraghavan, Vidyalakshmi Sethunath, Laura Heiser, Nicholas Wang, Charlotte K.Y. Ng, Edward S. Chen, Alexander Renwick, Tao Wang, Sarmistha Nanda, Martin Shea, Tamika Mitchell, Mahitha Rajendran, Ian Waters, Daniel J. Zabransky & 14 others Kenneth L. Scott, Carolina Gutierrez, Chandandeep Nagi, Felipe C. Geyer, Gary C. Chamness, Ben H. Park, Chad A. Shaw, Susan G. Hilsenbeck, Mothaffar F. Rimawi, Joe Gray, Britta Weigelt, Jorge S. Reis-Filho, C. Kent Osborne, Rachel Schiff

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Purpose: Resistance to anti-HER2 therapies in HER2+ breast cancer can occur through activation of alternative survival pathways or reactivation of the HER signaling network. Here we employed BT474 parental and treatment-resistant cell line models to investigate a mechanism by which HER2+ breast cancer can reactivate the HER network under potent HER2-targeted therapies. Experimental Design: Resistant derivatives to lapatinib (L), trastuzumab (T), or the combination (LR/TR/LTR) were developed independently from two independent estrogen receptor ER+/HER2+ BT474 cell lines (AZ/ATCC). Two derivatives resistant to the lapatinib-containing regimens (BT474/AZ-LR and BT474/ATCC-LTR lines) that showed HER2 reactivation at the time of resistance were subjected to massive parallel sequencing and compared with parental lines. Ectopic expression and mutant-specific siRNA interference were applied to analyze the mutation functionally. In vitro and in vivo experiments were performed to test alternative therapies for mutant HER2 inhibition. Results: Genomic analyses revealed that the HER2L755S mutation was the only common somatic mutation gained in the BT474/AZ-LR and BT474/ATCC-LTR lines. Ectopic expression of HER2L755S induced acquired lapatinib resistance in the BT474/AZ, SK-BR-3, and AU565 parental cell lines. HER2L755S-specific siRNA knockdown reversed the resistance in BT474/AZ-LR and BT474/ATCC-LTR lines. The HER1/2irreversible inhibitors afatinib and neratinib substantially inhibited both resistant cell growth and the HER2 and downstream AKT/MAPK signaling driven by HER2L755S in vitro and in vivo. Conclusions: HER2 reactivation through acquisition of the HER2L755S mutation was identified as a mechanism of acquired resistance to lapatinib-containing HER2-targeted therapy in preclinical HER2-amplified breast cancer models, which can be overcome by irreversible HER1/2 inhibitors.

Original languageEnglish (US)
Pages (from-to)5123-5134
Number of pages12
JournalClinical Cancer Research
Volume23
Issue number17
DOIs
StatePublished - Sep 1 2017

Fingerprint

Breast Neoplasms
Mutation
Cell Line
Small Interfering RNA
Therapeutics
Complementary Therapies
Estrogen Receptors
Research Design
lapatinib
Growth
In Vitro Techniques
Ectopic Gene Expression
BIBW 2992
Trastuzumab
N-(4-(3-chloro-4-(2-pyridinylmethoxy)anilino)-3-cyano-7-ethoxy-6-quinolyl)-4-(dimethylamino)-2-butenamide

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

HER2 reactivation through acquisition of the HER2 L755S mutation as a mechanism of acquired resistance to HER2-targeted therapy in HER2+ breast cancer. / Xu, Xiaowei; De Angelis, Carmine; Burke, Kathleen A.; Nardone, Agostina; Hu, Huizhong; Qin, Lanfang; Veeraraghavan, Jamunarani; Sethunath, Vidyalakshmi; Heiser, Laura; Wang, Nicholas; Ng, Charlotte K.Y.; Chen, Edward S.; Renwick, Alexander; Wang, Tao; Nanda, Sarmistha; Shea, Martin; Mitchell, Tamika; Rajendran, Mahitha; Waters, Ian; Zabransky, Daniel J.; Scott, Kenneth L.; Gutierrez, Carolina; Nagi, Chandandeep; Geyer, Felipe C.; Chamness, Gary C.; Park, Ben H.; Shaw, Chad A.; Hilsenbeck, Susan G.; Rimawi, Mothaffar F.; Gray, Joe; Weigelt, Britta; Reis-Filho, Jorge S.; Osborne, C. Kent; Schiff, Rachel.

In: Clinical Cancer Research, Vol. 23, No. 17, 01.09.2017, p. 5123-5134.

Research output: Contribution to journalArticle

Xu, X, De Angelis, C, Burke, KA, Nardone, A, Hu, H, Qin, L, Veeraraghavan, J, Sethunath, V, Heiser, L, Wang, N, Ng, CKY, Chen, ES, Renwick, A, Wang, T, Nanda, S, Shea, M, Mitchell, T, Rajendran, M, Waters, I, Zabransky, DJ, Scott, KL, Gutierrez, C, Nagi, C, Geyer, FC, Chamness, GC, Park, BH, Shaw, CA, Hilsenbeck, SG, Rimawi, MF, Gray, J, Weigelt, B, Reis-Filho, JS, Osborne, CK & Schiff, R 2017, 'HER2 reactivation through acquisition of the HER2 L755S mutation as a mechanism of acquired resistance to HER2-targeted therapy in HER2+ breast cancer', Clinical Cancer Research, vol. 23, no. 17, pp. 5123-5134. https://doi.org/10.1158/1078-0432.CCR-16-2191
Xu, Xiaowei ; De Angelis, Carmine ; Burke, Kathleen A. ; Nardone, Agostina ; Hu, Huizhong ; Qin, Lanfang ; Veeraraghavan, Jamunarani ; Sethunath, Vidyalakshmi ; Heiser, Laura ; Wang, Nicholas ; Ng, Charlotte K.Y. ; Chen, Edward S. ; Renwick, Alexander ; Wang, Tao ; Nanda, Sarmistha ; Shea, Martin ; Mitchell, Tamika ; Rajendran, Mahitha ; Waters, Ian ; Zabransky, Daniel J. ; Scott, Kenneth L. ; Gutierrez, Carolina ; Nagi, Chandandeep ; Geyer, Felipe C. ; Chamness, Gary C. ; Park, Ben H. ; Shaw, Chad A. ; Hilsenbeck, Susan G. ; Rimawi, Mothaffar F. ; Gray, Joe ; Weigelt, Britta ; Reis-Filho, Jorge S. ; Osborne, C. Kent ; Schiff, Rachel. / HER2 reactivation through acquisition of the HER2 L755S mutation as a mechanism of acquired resistance to HER2-targeted therapy in HER2+ breast cancer. In: Clinical Cancer Research. 2017 ; Vol. 23, No. 17. pp. 5123-5134.
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title = "HER2 reactivation through acquisition of the HER2 L755S mutation as a mechanism of acquired resistance to HER2-targeted therapy in HER2+ breast cancer",
abstract = "Purpose: Resistance to anti-HER2 therapies in HER2+ breast cancer can occur through activation of alternative survival pathways or reactivation of the HER signaling network. Here we employed BT474 parental and treatment-resistant cell line models to investigate a mechanism by which HER2+ breast cancer can reactivate the HER network under potent HER2-targeted therapies. Experimental Design: Resistant derivatives to lapatinib (L), trastuzumab (T), or the combination (LR/TR/LTR) were developed independently from two independent estrogen receptor ER+/HER2+ BT474 cell lines (AZ/ATCC). Two derivatives resistant to the lapatinib-containing regimens (BT474/AZ-LR and BT474/ATCC-LTR lines) that showed HER2 reactivation at the time of resistance were subjected to massive parallel sequencing and compared with parental lines. Ectopic expression and mutant-specific siRNA interference were applied to analyze the mutation functionally. In vitro and in vivo experiments were performed to test alternative therapies for mutant HER2 inhibition. Results: Genomic analyses revealed that the HER2L755S mutation was the only common somatic mutation gained in the BT474/AZ-LR and BT474/ATCC-LTR lines. Ectopic expression of HER2L755S induced acquired lapatinib resistance in the BT474/AZ, SK-BR-3, and AU565 parental cell lines. HER2L755S-specific siRNA knockdown reversed the resistance in BT474/AZ-LR and BT474/ATCC-LTR lines. The HER1/2irreversible inhibitors afatinib and neratinib substantially inhibited both resistant cell growth and the HER2 and downstream AKT/MAPK signaling driven by HER2L755S in vitro and in vivo. Conclusions: HER2 reactivation through acquisition of the HER2L755S mutation was identified as a mechanism of acquired resistance to lapatinib-containing HER2-targeted therapy in preclinical HER2-amplified breast cancer models, which can be overcome by irreversible HER1/2 inhibitors.",
author = "Xiaowei Xu and {De Angelis}, Carmine and Burke, {Kathleen A.} and Agostina Nardone and Huizhong Hu and Lanfang Qin and Jamunarani Veeraraghavan and Vidyalakshmi Sethunath and Laura Heiser and Nicholas Wang and Ng, {Charlotte K.Y.} and Chen, {Edward S.} and Alexander Renwick and Tao Wang and Sarmistha Nanda and Martin Shea and Tamika Mitchell and Mahitha Rajendran and Ian Waters and Zabransky, {Daniel J.} and Scott, {Kenneth L.} and Carolina Gutierrez and Chandandeep Nagi and Geyer, {Felipe C.} and Chamness, {Gary C.} and Park, {Ben H.} and Shaw, {Chad A.} and Hilsenbeck, {Susan G.} and Rimawi, {Mothaffar F.} and Joe Gray and Britta Weigelt and Reis-Filho, {Jorge S.} and Osborne, {C. Kent} and Rachel Schiff",
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TY - JOUR

T1 - HER2 reactivation through acquisition of the HER2 L755S mutation as a mechanism of acquired resistance to HER2-targeted therapy in HER2+ breast cancer

AU - Xu, Xiaowei

AU - De Angelis, Carmine

AU - Burke, Kathleen A.

AU - Nardone, Agostina

AU - Hu, Huizhong

AU - Qin, Lanfang

AU - Veeraraghavan, Jamunarani

AU - Sethunath, Vidyalakshmi

AU - Heiser, Laura

AU - Wang, Nicholas

AU - Ng, Charlotte K.Y.

AU - Chen, Edward S.

AU - Renwick, Alexander

AU - Wang, Tao

AU - Nanda, Sarmistha

AU - Shea, Martin

AU - Mitchell, Tamika

AU - Rajendran, Mahitha

AU - Waters, Ian

AU - Zabransky, Daniel J.

AU - Scott, Kenneth L.

AU - Gutierrez, Carolina

AU - Nagi, Chandandeep

AU - Geyer, Felipe C.

AU - Chamness, Gary C.

AU - Park, Ben H.

AU - Shaw, Chad A.

AU - Hilsenbeck, Susan G.

AU - Rimawi, Mothaffar F.

AU - Gray, Joe

AU - Weigelt, Britta

AU - Reis-Filho, Jorge S.

AU - Osborne, C. Kent

AU - Schiff, Rachel

PY - 2017/9/1

Y1 - 2017/9/1

N2 - Purpose: Resistance to anti-HER2 therapies in HER2+ breast cancer can occur through activation of alternative survival pathways or reactivation of the HER signaling network. Here we employed BT474 parental and treatment-resistant cell line models to investigate a mechanism by which HER2+ breast cancer can reactivate the HER network under potent HER2-targeted therapies. Experimental Design: Resistant derivatives to lapatinib (L), trastuzumab (T), or the combination (LR/TR/LTR) were developed independently from two independent estrogen receptor ER+/HER2+ BT474 cell lines (AZ/ATCC). Two derivatives resistant to the lapatinib-containing regimens (BT474/AZ-LR and BT474/ATCC-LTR lines) that showed HER2 reactivation at the time of resistance were subjected to massive parallel sequencing and compared with parental lines. Ectopic expression and mutant-specific siRNA interference were applied to analyze the mutation functionally. In vitro and in vivo experiments were performed to test alternative therapies for mutant HER2 inhibition. Results: Genomic analyses revealed that the HER2L755S mutation was the only common somatic mutation gained in the BT474/AZ-LR and BT474/ATCC-LTR lines. Ectopic expression of HER2L755S induced acquired lapatinib resistance in the BT474/AZ, SK-BR-3, and AU565 parental cell lines. HER2L755S-specific siRNA knockdown reversed the resistance in BT474/AZ-LR and BT474/ATCC-LTR lines. The HER1/2irreversible inhibitors afatinib and neratinib substantially inhibited both resistant cell growth and the HER2 and downstream AKT/MAPK signaling driven by HER2L755S in vitro and in vivo. Conclusions: HER2 reactivation through acquisition of the HER2L755S mutation was identified as a mechanism of acquired resistance to lapatinib-containing HER2-targeted therapy in preclinical HER2-amplified breast cancer models, which can be overcome by irreversible HER1/2 inhibitors.

AB - Purpose: Resistance to anti-HER2 therapies in HER2+ breast cancer can occur through activation of alternative survival pathways or reactivation of the HER signaling network. Here we employed BT474 parental and treatment-resistant cell line models to investigate a mechanism by which HER2+ breast cancer can reactivate the HER network under potent HER2-targeted therapies. Experimental Design: Resistant derivatives to lapatinib (L), trastuzumab (T), or the combination (LR/TR/LTR) were developed independently from two independent estrogen receptor ER+/HER2+ BT474 cell lines (AZ/ATCC). Two derivatives resistant to the lapatinib-containing regimens (BT474/AZ-LR and BT474/ATCC-LTR lines) that showed HER2 reactivation at the time of resistance were subjected to massive parallel sequencing and compared with parental lines. Ectopic expression and mutant-specific siRNA interference were applied to analyze the mutation functionally. In vitro and in vivo experiments were performed to test alternative therapies for mutant HER2 inhibition. Results: Genomic analyses revealed that the HER2L755S mutation was the only common somatic mutation gained in the BT474/AZ-LR and BT474/ATCC-LTR lines. Ectopic expression of HER2L755S induced acquired lapatinib resistance in the BT474/AZ, SK-BR-3, and AU565 parental cell lines. HER2L755S-specific siRNA knockdown reversed the resistance in BT474/AZ-LR and BT474/ATCC-LTR lines. The HER1/2irreversible inhibitors afatinib and neratinib substantially inhibited both resistant cell growth and the HER2 and downstream AKT/MAPK signaling driven by HER2L755S in vitro and in vivo. Conclusions: HER2 reactivation through acquisition of the HER2L755S mutation was identified as a mechanism of acquired resistance to lapatinib-containing HER2-targeted therapy in preclinical HER2-amplified breast cancer models, which can be overcome by irreversible HER1/2 inhibitors.

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DO - 10.1158/1078-0432.CCR-16-2191

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JO - Clinical Cancer Research

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