title: Correlation between messenger RNA expression and protein expression of immune checkpoint-associated molecules in bladder urothelial carcinoma: A retrospective study.
abstract: Immunotherapy for bladder cancer seems to have promising results. Here, we evaluated the association between messenger RNA (mRNA) and protein levels and possible prognostic value of the programmed cell death 1/programmed cell death ligand 1 (PD-1/PD-L1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) immune checkpoint pathways during bladder carcinogenesis.
  
  
  
  Tumor samples were obtained from 155 patients (84 with muscle-invasive bladder cancer [MIBC], and 71 non-muscle-invasive bladder cancer [NMIBC]) and normal bladder tissue from 15 patients. We evaluated the mRNA expression of 3 genes in the PD-1 pathway (PD-1, PD-L1, and PD-L2) and 4 in the CTLA4 pathway (CTLA4, CD28, CD80, and CD86) in normal and tumoral human bladder samples by quantitative real-time reverse transcription polymerase chain reaction, with immunohistochemistry used to evaluate the protein expression of PD-1 and PD-L1 in tumor and immune cells. Results of molecular analyses were compared with survival analyses.
  
  
  
  As compared with normal bladder tissue, MIBC tissue showed PD-1, PD-L1, CTLA4, and CD80 overexpression (59.5%, 60.7%, 84.5%, and 92.9%, respectively), whereas overexpression was lower in NMIBC tissue (22.5%, 4.2%, 35.2%, and 46.5%, respectively). The results of reverse transcription polymerase chain reaction analysis were confirmed by immunohistochemistry, with a high correlation between mRNA and protein expression. On multivariate analyses, overexpression of the studied genes was not associated with prognosis in relapse or progression of NMIBC or in recurrence-free and overall survival of MIBC.
  
  
  
  The CTLA4 pathway appears to be deregulated along with the PD-1/PD-L1 pathway in bladder carcinogenesis, with good correlation between mRNA and protein expression endorsing the useful role of immune checkpoints, especially for a large subgroup of MIBC.
authors.affiliation: Unité de Pharmacogénomique, Service de génétique, Institut Curie, Paris, France.
authors.forename: Constance
authors.initials: C
authors.lastname: Le Goux
authors.affiliation: Service d׳Anatomopathologie, Hôpital Cochin, Université Paris Descartes, Paris, France; Cancer et Immunité anti tumorale, INSERM U1138, Centre de Recherche des Cordeliers, Paris, France.
authors.forename: Diane
authors.initials: D
authors.lastname: Damotte
authors.affiliation: Unité de Pharmacogénomique, Service de génétique, Institut Curie, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Vacher
authors.affiliation: Service d׳Anatomopathologie, Hôpital Cochin, Université Paris Descartes, Paris, France.
authors.forename: Mathilde
authors.initials: M
authors.lastname: Sibony
authors.affiliation: Service d׳Urologie, Hôpital Cochin, Université Paris Descartes, Paris, France.
authors.forename: Nicolas Barry
authors.initials: NB
authors.lastname: Delongchamps
authors.affiliation: Unité de Pharmacogénomique, Service de génétique, Institut Curie, Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Schnitzler
authors.affiliation: Service d׳Anatomopathologie, Hôpital Cochin, Université Paris Descartes, Paris, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Terris
authors.affiliation: Service d׳Urologie, Hôpital Cochin, Université Paris Descartes, Paris, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Zerbib
authors.affiliation: Unité de Pharmacogénomique, Service de génétique, Institut Curie, Paris, France; Université Paris Descartes, Paris, France.
authors.forename: Ivan
authors.initials: I
authors.lastname: Bieche
authors.affiliation: Service d׳Urologie, Institut Paoli-Calmettes, Marseille, France. Electronic address: pignotg@ipc.unicancer.fr.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Pignot
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018122:B7-1 Antigen / Q000235:genetics / Q000378:metabolism; D051940:B7-2 Antigen / Q000235:genetics / Q000378:metabolism; D060890:B7-H1 Antigen / Q000235:genetics / Q000378:metabolism; D018106:CD28 Antigens / Q000235:genetics / Q000378:metabolism; D060908:CTLA-4 Antigen / Q000235:genetics / Q000378:metabolism; D002295:Carcinoma, Transitional Cell / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D061027:Programmed Cell Death 1 Ligand 2 Protein / Q000235:genetics; D061026:Programmed Cell Death 1 Receptor / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012189:Retrospective Studies; D015398:Signal Transduction; D001743:Urinary Bladder / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology
chemical_list: D018122:B7-1 Antigen; D051940:B7-2 Antigen; D060890:B7-H1 Antigen; C423236:CD274 protein, human; D018106:CD28 Antigens; C497964:CD86 protein, human; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C105992:PDCD1 protein, human; D061027:Programmed Cell Death 1 Ligand 2 Protein; D061026:Programmed Cell Death 1 Receptor; D012333:RNA, Messenger
keywords: Bladder cancer; CTLA4; Immune checkpoints; Immunohistochemistry; Molecular marker; PD-1; RT-PCR