Author Manuscript Published OnlineFirst on April 15, 2019; DOI: 10.1158/1078-0432.CCR-19-0160 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Title: Detection of NRG1 gene fusions in solid tumors Authors: Sushma Jonna1, Rebecca A. Feldman2, Jeffrey Swensen2, Zoran Gatalica2, Wolfgang M. Korn2, Hossein Borghaei3, Patrick C. Ma4, Jorge Nieva5, Alexander Spira6, Ari Vanderwalde7, Antoinette Wozniak8, Edward S. Kim9, Stephen V. Liu1 Affiliations: 1 Georgetown University, Washington, DC 2 Caris Life Sciences, Phoenix, AZ 3 Fox Chase Cancer Center, Philadelphia, PA 4 WVU Cancer Institute, West Virginia University, Morgantown, WV 5 University of Southern California, Los Angeles, CA 6 Virginia Health Specialists, Fairfax, VA 7 West Cancer Center, Memphis, TN 8 University of Pittsburgh Medical Center, Pittsburgh, PA 9 Atrium Healthcare, Levine Cancer Institute, Charlotte, NC Running Title: NRG1 fusions in solid tumors Keywords: NRG1, Neuregulin-1, gene fusions, NSCLC, RNA-sequencing Financial Support: The authors received no specific funding for this work. Corresponding Author: Stephen V. Liu, MD Georgetown University Lombardi Comprehensive Cancer Center 3800 Reservoir Road NW Washington, DC 20007 Phone: 202-444-2223 Fax: 202-444-7009 Email: [email protected] Conflict of Interest Statement: S. Jonna reports no competing interests. R.A. Feldman, J. Swensen, Z. Gatalica and W.M. Korn are employees of Caris Life Sciences. W.M. Korn reports consulting/advisory role from Merrimack and Merck Sharp & Dohme, stock/ownership interests with OncoCyte, has received honoraria from Genentech/Roche and institution-associated funding from Merrimack. H. Borghaei reports consulting/advisory role from Bristol-Myers Squibb, Lilly, Celgene, Genentech, Pfizer, Boehringer Ingelheim, EMD Serono, Trovagene, Novartis, Merck, AstraZeneca, Genmab, Regeneron, Contargia AB, BioNTech AG, Abbvie, received honoraria from Bristol-Myers Squibb, Celgene, Axiom Biotechnologies and institution-associated research funding from Millennium, Merck, Celgene, Bristol-Myers Squibb and Lilly. P.C. Ma reports association with the speakers’ bureau for Merck, Takeda, Bristol-Myers Squibb, stock/ownership from Cymeta Biopharmaceutical and institution-associated research funding from Pfizer, MedImmune, AstraZeneca, Bristol-Myers Squibb, Loxo, 1 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on April 15, 2019; DOI: 10.1158/1078-0432.CCR-19-0160 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Xcovery, Abbvie, EpicentRx, Tesaro, Boehringer Ingelheim, OncoMed, CBT Pharmaceuticals, Merck and Cymeta Biopharmaceutical. J. Nieva reports consulting/advisory role with AstraZeneca, Genentech/Roche and Western Oncolytics, stock/ownership in Epic Sciences and research funding from Merck. A. Spira reports institution-associated research funding from Roche, AstraZeneca, Boehringer Ingelheim, Astellas Pharma, MedImmune, Novartis, Newlink Genetics, Incyte, Abbvie, Ignyta, LAM Therapeutics, Trovagene, Takeda, Macrogenics and CytomX Therapeutics. A. Vanderwalde reports consulting/advisory role with Bristol-Myers Squibb, AstraZeneca, Genentech and Caris Life Sciences, has received honoraria from AstraZeneca and institution-associated research funding from Amgen, Merck, Genentech/Roche, Millennium, AstraZeneca, Polynoma, Lilly, Bristol-Myers Squibb and Amgen. A. Wozniak reports consulting/advisory role with Boehringer Ingelheim, AstraZeneca, ARIAD, Coherus Biosciences, Array BioPharma, Hospira, Quintiles, BeyondSpring Pharmaceuticals, WCCT Global, Epic, Huron Consulting, HUYA Bioscience International and Takeda and has received research funding from Boehringer Ingelheim. E.S. Kim reports consulting/advisory role with Lilly, AstraZeneca, Boehringer Ingelheim, Pfizer, Merck and Takeda, has received honoraria from Lilly, AstraZeneca, Boehringer Ingelheim, Pfizer, Merck and Takeda and research funding from Boehringer Ingelheim, Lilly, Merck, Ignyta and Genentech/Roche. S.V. Liu reports consulting/advisory role with Apollomics, Boehringer-Ingelheim, G1 Therapeutics, Genentech/Roche, Ignyta, Guardant 360, Inivata, Janssen, Pfizer, Lilly, Merck, Taiho Pharmaceutical, Bristol-Myers Squibb, AstraZeneca, Takeda, HERON and Regeneron and has received research funding from Genentech/Roche, Pfizer, Threshold Pharmaceuticals, Clovis Oncology, Corvus Pharmaceuticals, Esanex, Bayer, OncoMed, Merck, Lycera, AstraZeneca, Ignyta, Molecular Partners, Blueprint Medicines, Lilly and Rain Therapeutics. Word Count: 2,381 Figure Count: 8 Table Count: 2 2 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on April 15, 2019; DOI: 10.1158/1078-0432.CCR-19-0160 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Significance: NRG1 fusions are potentially actionable genomic events seen in various tumor types. While there are reports of therapeutic efficacy with agents that target Erb-B2/Erb-B3, little is known about the characteristics of these fusions. Here, we report the incidence of NRG1 fusions in a large cohort of solid tumors that underwent RNA sequencing. NRG1 fusions were detected at a low incidence across many solid tumor types. Multiple fusion partners were identified, which will influence the development of strategies to detect these events on a large scale. 3 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on April 15, 2019; DOI: 10.1158/1078-0432.CCR-19-0160 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Abstract: Background: NRG1 gene fusions are rare but potentially actionable oncogenic drivers present in some solid tumors. Details regarding the incidence of these gene rearrangements are lacking. Here, we assessed the incidence of NRG1 fusions across multiple tumor types and described fusion partners. Methods: Tumor specimens submitted for molecular profiling at a CLIA-certified genomics laboratory and that underwent fusion testing by anchored multiplex PCR for targeted RNA sequencing were retrospectively identified. The overall and tumor-specific incidence was noted as was the specific fusion partner. Results: Out of 21,858 tumor specimens profiled from September 2015 to December 2018, 41 cases (0.2%) harbored an NRG1 fusion. Multiple fusion partners were identified. Fusion events were seen across tumor types. The greatest incidence was in non-small cell lung cancer (25), though this represented only 0.3% of non-small cell lung cancer cases tested. Other tumor types harboring an NRG1 fusion included gallbladder cancer, renal cell carcinoma, bladder cancer, ovarian cancer, pancreatic cancer, breast cancer, neuroendocrine tumor, sarcoma, and colorectal cancer. Conclusion: NRG1 fusions can be detected at a low incidence across multiple tumor types with significant heterogeneity in fusion partner. 4 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on April 15, 2019; DOI: 10.1158/1078-0432.CCR-19-0160 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Introduction: Appropriate management of advanced non-small cell lung cancer (NSCLC) is guided by the presence or absence of specific molecular drivers. The identification of activating genomic alterations in EGFR, ALK, ROS1 or BRAF not only provides insight into the underlying biology but also directs initial and subsequent therapeutic decisions (1–5). It is now standard of care to search for these mutations and fusions in all patients with non-squamous NSCLC (6). It has also become clear that some molecular drivers will serve as therapeutic targets across multiple tumor types (7), including the tumor agnostic approval of larotrectinib for tumors with a gene fusion in NTRK1, NTRK2, or NTRK3 (8). As our understanding of cancer grows increasingly sophisticated, additional drivers have surfaced that may have a similar impact on evolving treatment paradigms. Neuregulin-1 (NRG1) gene fusions are an emerging, potentially actionable oncogenic driver (9). NRG1 fusions can promote pathologic signaling via MAPK and other canonical pathways (10). When NRG1 fusions are present, targeting ERBB2 and ERBB3 has been an effective treatment strategy in vitro. Recently, clinical responses to tyrosine kinase inhibitors and monoclonal antibodies have also been reported (9,11– 13). The interest in evaluating the prevalence of NRG1 fusions has increased given the potential therapeutic implications of this genetic alteration. Since the original description of the CD74-NRG1 gene fusion in invasive mucinous lung adenocarcinoma, detection has been noted in other tumor types, both de novo and as a resistance mechanism in ALK rearranged NSCLC (9,14–16). Here, we report the incidence and characteristics of 5 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on April 15, 2019; DOI: 10.1158/1078-0432.CCR-19-0160 Author manuscripts have been peer reviewed and accepted for publication but have not yet