BACKGROUND: The molecular content of urine is defined by filtration in the kidneys and by local release from tissues lining the urinary tract. Pathological processes and different therapies change the molecular composition of urine and a variety of markers have been analyzed in patients with bladder cancer. The response to BCG immunotherapy and chemotherapy has been extensively studied and elevated urine concentrations of IL-1RA, IFN-α, IFN-γ TNF-α, and IL-17 have been associated with improved outcome. METHODS: In this study, the host response to intravesical alpha 1-oleate treatment was characterized in patients with non-muscle invasive bladder cancer by proteomic and transcriptomic analysis. RESULTS: Proteomic profiling detected a significant increase in multiple cytokines in the treatment group compared to placebo. The innate immune response was strongly activated, including IL-1RA and pro-inflammatory cytokines in the IL-1 family (IL-1α, IL-1β, IL-33), chemokines (MIP-1α, IL-8), and interferons (IFN-α2, IFN-γ). Adaptive immune mediators included IL-12, Granzyme B, CD40, PD-L1, and IL-17D, suggesting broad effects of alpha 1-oleate treatment on the tumor tissues. CONCLUSIONS: The cytokine response profile in alpha 1-oleate treated patients was similar to that reported in BCG treated patients, suggesting a significant overlap. A reduction in protein levels at the end of treatment coincided with inhibition of cancer-related gene expression in tissue biopsies, consistent with a positive treatment effect. Thus, in addition to killing tumor cells and inducing cell detachment, alpha 1-oleate is shown to activate a broad immune response with a protective potential.

Department of Biomedical Engineering University of Houston Houston Texas USA

Department of Pathology and Molecular Medicine Motol University Hospital 2nd Faculty of Medicine Charles University Praha Prague Czech Republic

Department of Urology Motol University Hospital 2nd Faculty of Medicine Charles University Praha Prague Czech Republic

Division of Microbiology Immunology and Glycobiology Department of Laboratory Medicine Faculty of Medicine Lund University Lund Sweden

Zobrazit více v PubMed

Antoni S, Ferlay J, Soerjomataram I, Znaor A, Jemal A, Bray F. Bladder cancer incidence and mortality: a global overview and recent trends. Eur Urol. 2017;71:96‐108. PubMed

Torre LA, Bray F, Siegel RL, Ferlay J, Lortet‐Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87‐108. PubMed

van Rhijn BWG, Burger M, Lotan Y, et al. Recurrence and progression of disease in non–muscle‐invasive bladder cancer: from epidemiology to treatment strategy. Eur Urol. 2009;56:430‐442. PubMed

Malats N, Real FX. Epidemiology of bladder cancer. Hematol/Oncol Clin. 2015;29:177‐189. PubMed

Hien TT, Ambite I, Butler D, et al. Bladder cancer therapy without toxicity—a dose‐escalation study of alpha1‐oleate. Int J Cancer. 2020;147:2479‐2492. PubMed

Brisuda A, Ho JCS, Kandiyal PS, et al. Bladder cancer therapy using a conformationally fluid tumoricidal peptide complex. Nat Commun. 2021;12:3427. PubMed PMC

Frantzi M, Vlahou A. Ten years of proteomics in bladder cancer: progress and future directions. Bladder Cancer. 2017;3:1‐18. PubMed PMC

Santoni G, Morelli MB, Amantini C, Battelli N. Urinary markers in bladder cancer: an update. Front Oncol. 2018;8:362. PubMed PMC

Duquesne I, Weisbach L, Aziz A, Kluth LA, Xylinas E. The contemporary role and impact of urine‐based biomarkers in bladder cancer. Transl Androl Urol. 2017;6:1031‐1042. PubMed PMC

Sugeeta SS, Sharma A, Ng K, Nayak A, Vasdev N. Biomarkers in bladder cancer surveillance. Front Surg. 2021;8:735868. PubMed PMC

Barak V, Itzkovich D, Einarsson R, Gofrit O, Pode D. Non‐invasive detection of bladder cancer by UBC rapid test, ultrasonography and cytology. Anticancer Res. 2020;40:3967‐3972. PubMed

Chakraborty A, Dasari S, Long W, Mohan C. Urine protein biomarkers for the detection, surveillance, and treatment response prediction of bladder cancer. Am J Cancer Res. 2019;9:1104‐1117. PubMed PMC

De Boer EC, De Jong WH, Steerenberg PA, et al. Induction of urinary interleukin‐1 (IL‐1), IL‐2, IL‐6, and tumour necrosis factor during intravesical immunotherapy with bacillus Calmette‐Guérin in superficial bladder cancer. Cancer Immunol Immunother. 1992;34:306‐312. PubMed PMC

Witjes JA, APMvd M, Collette L, et al. Long‐term follow‐up of an EORTC randomized prospective trial comparing intravesical bacille calmette–guérin–RIVM and mitomycin c in superficial bladder cancer. Urology. 1998;52:403‐410. PubMed

de Boer EC, Somogyi L, de Ruiter GJW, de Reijke TM, Kurth KH, Schamhart DHJ. Role of interleukin‐8 in onset of the immune response in intravesical BCG therapy for superficial bladder cancer. Urol Res. 1997;25:31‐34. PubMed

Kawai K, Miyazaki J, Joraku A, Nishiyama H, Akaza H. BacillusCalmette–Guerin (BCG) immunotherapy for bladder cancer: current understanding and perspectives on engineeredBCGvaccine. Cancer Sci. 2013;104:22‐27. PubMed PMC

Kamat AM, Briggman J, Urbauer DL, et al. Cytokine panel for response to intravesical therapy (CyPRIT): nomogram of changes in urinary cytokine levels predicts patient response to Bacillus Calmette–Guérin. Eur Urol. 2016;69:197‐200. PubMed PMC

Schneider T, Zhao L, Zhu Z, et al. The past, present, and future of immunotherapy for bladder tumors. Med Oncol. 2022;39:236. PubMed

Papageorgiou A, Dinney CPN, McConkey DJ. Interferon‐α induces TRAIL expression and cell death via an IRF‐1‐dependent mechanism in human bladder cancer cells. Cancer Biol Ther. 2007;6:872‐879. PubMed

Brausi M, Oddens J, Sylvester R, et al. Side effects of Bacillus Calmette–Guérin (BCG) in the treatment of intermediate‐ and high‐risk Ta, T1 papillary carcinoma of the bladder: results of the EORTC genito‐urinary cancers group randomised phase 3 study comparing one‐third dose with full dose and 1 year with 3 years of maintenance BCG. Eur Urol. 2014;65:69‐76. PubMed

Jiang S, Redelman‐Sidi G. BCG in bladder cancer immunotherapy. Cancers. 2022;14:3073. PubMed PMC

Gocher AM, Workman CJ, Vignali DAA. Interferon‐gamma: teammate or opponent in the tumour microenvironment? Nat Rev Immunol. 2022;22:158‐172. PubMed PMC

Mossberg AK, Hou Y, Svensson M, Holmqvist B, Svanborg C. HAMLET treatment delays bladder cancer development. J Urol. 2010;183:1590‐1597. PubMed

Pettenati C, Ingersoll MA. Mechanisms of BCG immunotherapy and its outlook for bladder cancer. Nat Rev Urol. 2018;15:615‐625. PubMed

Conti P, Reale M, Nicolai M, et al. Bacillus Calmette–Guérin potentiates monocyte responses to lipopolysaccharide‐induced tumor necrosis factor and interleukin‐1, but not interleukin‐6 in bladder cancer patients. Cancer Immunol Immunother. 1994;38:365‐371. PubMed PMC

Cai T, Nesi G, Mazzoli S, et al. Prediction of response to bacillus Calmette–Guérin treatment in non‐muscle invasive bladder cancer patients through interleukin‐6 and interleukin‐10 ratio. Exp Ther Med. 2012;4:459‐464. PubMed PMC

Nunez‐Nateras R, Castle EP, Protheroe CA, et al. Predicting response to bacillus Calmette–Guérin (BCG) in patients with carcinoma in situ of the bladder. Urol Oncol. 2014;32(1):e23‐e30. PubMed PMC

Zhang N, Jiang G, Liu X, Na R, Wang X, Xu J. Prediction of Bacillus Calmette–Guerin response in patients with bladder cancer after transurethral resection of bladder tumor by using genetic variation based on genomic studies. Biomed Res Int. 2016;2016:e9859021. PubMed PMC

Han J, Gu X, Li Y, Wu Q. Mechanisms of BCG in the treatment of bladder cancer‐current understanding and the prospect. Biomed Pharmacother. 2020;129:110393. PubMed

Sfakianos JP, Salome B, Daza J, Farkas A, Bhardwaj N, Horowitz A. Bacillus Calmette–Guerin (BCG): its fight against pathogens and cancer. Urol Oncol. 2021;39:121‐129. PubMed

Crispen PL, Kusmartsev S. Mechanisms of immune evasion in bladder cancer. Cancer Immunol Immunother. 2020;69:3‐14. PubMed PMC

John A, Günes C, Bolenz C, et al. Bladder cancer‐derived interleukin‐1 converts the vascular endothelium into a pro‐inflammatory and pro‐coagulatory surface. BMC Cancer. 2020;20:1178. PubMed PMC

Jonasch E, Haluska FG. Interferon in oncological practice: review of interferon biology, clinical applications, and toxicities. Oncologist. 2001;6:34‐55. PubMed

Kamat AM, Lamm DL. Immunotherapy for bladder cancer. Curr Urol Rep. 2001;2:62‐69. PubMed

Marth C, Fiegl H, Zeimet AG, et al. Interferon‐γ expression is an independent prognostic factor in ovarian cancer. Am J Obstet Gynecol. 2004;191:1598‐1605. PubMed

Papageorgiou A, Lashinger L, Millikan R, et al. Role of tumor necrosis factor‐related apoptosis‐inducing ligand in interferon‐induced apoptosis in human bladder cancer cells. Cancer Res. 2004;64:8973‐8979. PubMed

Goulet CR, Champagne A, Bernard G, et al. Cancer‐associated fibroblasts induce epithelial–mesenchymal transition of bladder cancer cells through paracrine IL‐6 signalling. BMC Cancer. 2019;19:137. PubMed PMC

Nilsson MB, Langley RR, Fidler IJ. Interleukin‐6, secreted by human ovarian carcinoma cells, is a potent proangiogenic cytokine. Cancer Res. 2005;65:10794‐10800. PubMed PMC

Yao JS, Zhai W, Young WL, Yang GY. Interleukin‐6 triggers human cerebral endothelial cells proliferation and migration: the role for KDR and MMP‐9. Biochem Biophys Res Commun. 2006;342:1396‐1404. PubMed

Henn V, Slupsky JR, Gräfe M, et al. CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells. Nature. 1998;391:591‐594. PubMed

Chatzigeorgiou A, Lyberi M, Chatzilymperis G, Nezos A, Kamper E. CD40/CD40L signaling and its implication in health and disease. Biofactors. 2009;35:474‐483. PubMed

Munroe ME, Bishop GA. A costimulatory function for T cell CD401. J Immunol. 2007;178:671‐682. PubMed

Burke JD, Young HA. IFN‐γ: a cytokine at the right time, is in the right place. Semin Immunol. 2019;43:101280. PubMed PMC

Ambite I, Butler D, Wan MLY, et al. Molecular determinants of disease severity in urinary tract infection. Nat Rev Urol. 2021;18:468‐486. PubMed PMC

Butler D, Ambite I, Wan MLY, Tran TH, Wullt B, Svanborg C. Immunomodulation therapy offers new molecular strategies to treat UTI. Nat Rev Urol. 2022;19:419‐437. PubMed PMC

Wullt B, Butler DSC, Ambite I, Kinsolving J, Krintel C, & Svanborg C. Immunomodulation—a molecular solution to treating patients with severe bladder pain syndrome? European Urology Open Science. 2021;31:49‐58. PubMed PMC

Shore ND, Boorjian SA, Canter DJ, et al. Intravesical rAd–IFNα/Syn3 for patients with high‐grade, Bacillus Calmette–Guerin–refractory or relapsed non–muscle‐invasive bladder cancer: a phase II randomized study. JCO. 2017;35:3410‐3416. PubMed PMC

Maimela NR, Liu S, Zhang Y. Fates of CD8+ T cells in tumor microenvironment. Comput Struct Biotechnol J. 2019;17:1‐13. PubMed PMC

Ding H, Wang G, Yu Z, Sun H, Wang L. Role of interferon‐gamma (IFN‐gamma) and IFN‐gamma receptor 1/2 (IFNgammaR1/2) in regulation of immunity, infection, and cancer development: IFN‐gamma‐dependent or independent pathway. Biomed Pharmacother. 2022;155:113683. PubMed

Briesemeister D, Sommermeyer D, Loddenkemper C, et al. Tumor rejection by local interferon gamma induction in established tumors is associated with blood vessel destruction and necrosis. Int J Cancer. 2011;128:371‐378. PubMed

Droller MJ, Gomolka D. Enhancement of natural cytotoxicity in lymphocytes from animals with carcinogen‐induced bladder cancer. J Urol. 1983;129:625‐629. PubMed

Parronchi P, De Carli M, Manetti R, et al. IL‐4 and IFN (alpha and gamma) exert opposite regulatory effects on the development of cytolytic potential by Th1 or Th2 human T cell clones. J Immunol. 1992;149:2977‐2983. PubMed

Mojic M, Takeda K, Hayakawa Y. The dark side of IFN‐γ: its role in promoting cancer immunoevasion. Int J Mol Sci. 2018;19:89. PubMed PMC

Schonbeck U, Varo N, Libby P, Buring J, Ridker PM. Soluble CD40L and cardiovascular risk in women. Circulation. 2001;104:2266‐2268. PubMed

Giat E, Ehrenfeld M, Shoenfeld Y. Cancer and autoimmune diseases. Autoimmun Rev. 2017;16:1049‐1057. PubMed

Clinical and molecular response to alpha1-oleate treatment in patients with bladder cancer