{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T08:14:39Z","timestamp":1771056879717,"version":"3.50.1"},"posted":{"date-parts":[[2026,1,26]]},"group-title":"Cancer Biology","reference-count":29,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T00:00:00Z","timestamp":1769385600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH\/NCI","award":["P50CA091846"],"award-info":[{"award-number":["P50CA091846"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Purpose<\/jats:title>\n                  <jats:p>Intravesical interferon-alpha (IFN\u03b1) gene therapy has shown promise in treating BCG-unresponsive non-muscle invasive bladder cancer (NMIBC). Ongoing work in our lab aims to further improve its treatment efficacy by identifying resistance mechanisms and deploying targeted combination treatment strategies.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Experimental design<\/jats:title>\n                  <jats:p>\n                    We performed end-tumor RNA-seq analysis of MB49 murine tumors treated with IFN\u03b1 gene therapy, identifying the ErbB pathway as a resistance mechanism. We consequently hypothesized that a combination treatment involving an ErbB pathway blocker and IFN\u03b1 could yield improved outcomes. MB49 cells were treated\n                    <jats:italic>in vitro<\/jats:italic>\n                    with lentiviral IFN\u03b1 (LV-IFN\u03b1) gene therapy, with\/without Afatinib, a pan-ErbB inhibitor, and cell viability and migration assays were performed. Next,\n                    <jats:italic>in vivo<\/jats:italic>\n                    studies were conducted in the syngeneic MB49 orthotopic murine bladder cancer model. The mice were randomized into 5 treatment groups (n=10 each): saline (Ctrl), LV-Ctrl, oral Afatinib monotherapy, intravesical LV-IFN\u03b1 monotherapy, and the experimental intravesical LV-IFN\u03b1 + oral Afatinib combination therapy. Overall survival (OS) and drug toxicity were assessed.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Combination therapy significantly reduced MB49 cell viability\n                    <jats:italic>in vitro<\/jats:italic>\n                    compared to all other treatment conditions (mean relative ATPase activity at 72 h for the combination treatment was 4%, compared to 100%, 26%, and 28% for Ctrl, LV-IFN\u03b1, and Afatinib, respectively, p&lt;0.001). This additive effect on cell viability appeared to be driven by a combination of early-cytostatic and late-cytolytic effects. The combination treatment also markedly inhibited cell migration (mean migrated cells\/10x Boyden chamber assay at 36 h were: 92.3 for the combination therapy and 631.0, 600.4, and 270.3 for Ctrl, LV-IFN\u03b1, and Afatinib, respectively, p&lt;0.001). Finally, the\n                    <jats:italic>in vivo<\/jats:italic>\n                    studies demonstrated improved OS with combination therapy (median OS was 49 d in the combination group vs 15, 29, and 26 d in Ctrl, LV-IFN\u03b1, and Afatinib groups, respectively, Log-rank p&lt;0.001). No mice in the combination therapy group died of drug toxicity.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Our preliminary findings suggest that the ErbB pathway may serve as a clinically significant resistance mechanism to intravesical IFN\u03b1 gene therapy, and when targeted concurrently, may improve treatment efficacy.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.01.23.701130","type":"posted-content","created":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T23:35:11Z","timestamp":1769470511000},"source":"Crossref","is-referenced-by-count":0,"title":["Combination Treatment with Intravesical Interferon-Alpha Gene Therapy and Oral Pan-ErbB Receptor Family Blocker Improves Survival in Mice with Bladder Cancer"],"prefix":"10.64898","author":[{"given":"Akshay","family":"Sood","sequence":"first","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"},{"name":"Department of Urology, The James Cancer Hospital and Solove Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alberto","family":"Martini","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan K.","family":"Rudzinski","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Come","family":"Tholomier","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberto","family":"Contieri","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"I-Ling","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nigel R.","family":"Parker","sequence":"additional","affiliation":[{"name":"A.I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seppo","family":"Yla-Herttuala","sequence":"additional","affiliation":[{"name":"A.I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David J.","family":"McConkey","sequence":"additional","affiliation":[{"name":"Department of Urology, University of Rochester Medical Center, Rochester, NY"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Colin P.N.","family":"Dinney","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"},{"name":"Department of Urology, University of Rochester Medical Center, Rochester, NY"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0885-2369","authenticated-orcid":false,"given":"Sharada","family":"Mokkapati","sequence":"additional","affiliation":[{"name":"Department of Urology, The University of Texas MD Anderson Cancer Center, TX"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026021323551146000_2026.01.23.701130v2.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2008.12.010"},{"key":"2026021323551146000_2026.01.23.701130v2.2","doi-asserted-by":"publisher","DOI":"10.1038\/nrc1609"},{"key":"2026021323551146000_2026.01.23.701130v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.09.007"},{"key":"2026021323551146000_2026.01.23.701130v2.4","doi-asserted-by":"crossref","first-page":"3410","DOI":"10.1200\/JCO.2017.72.3064","article-title":"Intravesical rAd-IFNalpha\/Syn3 for Patients With High-Grade, Bacillus Calmette-Guerin-Refractory or Relapsed Non-Muscle-Invasive Bladder Cancer: A Phase II Randomized Study","volume":"35","year":"2017","journal-title":"J Clin Oncol"},{"key":"2026021323551146000_2026.01.23.701130v2.5","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/S1470-2045(20)30540-4","article-title":"Intravesical nadofaragene firadenovec gene therapy for BCG-unresponsive non-muscle-invasive bladder cancer: a single-arm, open-label, repeat-dose clinical trial","volume":"22","year":"2021","journal-title":"Lancet Oncol"},{"key":"2026021323551146000_2026.01.23.701130v2.6","doi-asserted-by":"publisher","DOI":"10.1097\/JU.0000000000004020"},{"key":"2026021323551146000_2026.01.23.701130v2.7","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.eururo.2021.12.009","article-title":"Antiadenovirus Antibodies Predict Response Durability to Nadofaragene Firadenovec Therapy in BCG-unresponsive Non-muscle-invasive Bladder Cancer: Secondary Analysis of a Phase 3 Clinical Trial","volume":"81","year":"2022","journal-title":"Eur Urol"},{"key":"2026021323551146000_2026.01.23.701130v2.8","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.omto.2022.06.005","article-title":"Lentiviral interferon: A novel method for gene therapy in bladder cancer","volume":"26","year":"2022","journal-title":"Mol Ther Oncolytics"},{"key":"2026021323551146000_2026.01.23.701130v2.9","doi-asserted-by":"publisher","DOI":"10.1038\/nrc3858"},{"key":"2026021323551146000_2026.01.23.701130v2.10","doi-asserted-by":"publisher","DOI":"10.1158\/1538-7445.AM2018-3954"},{"key":"2026021323551146000_2026.01.23.701130v2.11","doi-asserted-by":"crossref","unstructured":"Schneider, C. 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