American Association for Cancer Research
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Supplementary Figures from Substance P Autocrine Signaling Contributes to Persistent HER2 Activation That Drives Malignant Progression and Drug Resistance in Breast Cancer

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journal contribution
posted on 2023-03-30, 21:55 authored by Susana Garcia-Recio, Gemma Fuster, Patricia Fernandez-Nogueira, Eva M. Pastor-Arroyo, So Yeon Park, Cristina Mayordomo, Elisabet Ametller, Mario Mancino, Xavier Gonzalez-Farre, Hege G. Russnes, Pablo Engel, Domiziana Costamagna, Pedro L. Fernandez, Pedro Gascón, Vanessa Almendro

PDF file, 12332K, Supplementary Figure S1. Expression of NK-1R, NK-2R and SP in normal mammary epithelium. Supplementary Figure S2. Transactivation of HER2 by human tachykinins. Supplementary Figure S3. Dose-response effect of SP on HER2 activation. Supplementary Figure S4. Contribution of HER2 signaling to SP-induced MAPK activation. Supplementary Figure S5. Representative images of NK-1R immunohistochemistry in the tumors used for primary cultures. Supplementary Figure S6. Activation of HER2 under SP treatment in each primary culture derived from 19 primary breast tumors. Supplementary Figure S7. Overexpression of NK-1R in BC cells. Supplementary Figure S8. Mitotic index for the MDA-MB-231 and MDA-MB-453 xenograft tumors growth under the continuous exposure to SP. Supplementary Figure S9. Immunohistochemical analysis of the xenograft tumors treated with the NK-1R inhibitor L-733,060. Supplementary Figure S10. Acute and chronic effects of SP on cellular responses to anti-HER2 and anti-EGFR therapies. Supplementary Table S1. Statistical analysis for the correlation of NK-1R, NK-2R and SP expression levels. Supplementary Table S2. Statistical analysis for the correlation of NK-1R, NK-2R and SP expression levels. Supplementary Table S3. Clinical information of the cohort used to investigate the levels of circulating SP in healthy donors and breast cancer patients. Supplementary Table S4. Histopathologic information of the samples used Supplementary Table S5. IC50 obtained in the drug-response studies and combination index.



ERBB receptor transmodulation by heterologous G-protein–coupled receptors (GPCR) generates functional diversity in signal transduction. Tachykinins are neuropeptides and proinflammatory cytokines that promote cell survival and cancer progression by activating several GPCRs. In this work, we found that the pain-associated tachykinin Substance P (SP) contributes to persistent transmodulation of the ERBB receptors, EGFR and HER2, in breast cancer, acting to enhance malignancy and therapeutic resistance. SP and its high-affinity receptor NK-1R were highly expressed in HER2+ primary breast tumors (relative to the luminal and triple-negative subtypes) and were overall correlated with poor prognosis factors. In breast cancer cell lines and primary cultures derived from breast cancer samples, we found that SP could activate HER2. Conversely, RNA interference-mediated attenuation of NK-1R, or its chemical inhibition, or suppression of overall GPCR-mediated signaling, all strongly decreased steady-state expression of EGFR and HER2, establishing that their basal activity relied upon transdirectional activation by GPCR. Thus, SP exposure affected cellular responses to anti-ERBB therapies. Our work reveals an important oncogenic cooperation between NK-1R and HER2, thereby adding a novel link between inflammation and cancer progression that may be targetable by SP antagonists that have been clinically explored. Cancer Res; 73(21); 6424–34. ©2013 AACR.