RESUMO
Poor response to Bacillus Calmette-Guérin (BCG) immunotherapy remains a major barrier in the management of patients with non-muscle invasive bladder cancer (NMIBC). Multiple factors are associated with poor outcomes, including biological aging and female sex. More recently, it has emerged that a B-cell infiltrated pre-treatment immune microenvironment of NMIBC tumors can influence the response to intra-vesically administered BCG. The mechanisms underlying the roles of B cells in NMIBC are poorly understood. Here, we show that B-cell dominant tertiary lymphoid structures (TLSs), a hallmark feature of the chronic mucosal immune response, are abundant and located close to the epithelial compartment in pre-treatment tumors from BCG non-responders. Digital spatial proteomic profiling of whole tumor sections from male and female patients with NMIBC who underwent treatment with intravesical BCG, revealed higher expression of immune exhaustion-associated proteins within the tumor-adjacent TLSs in both responders and non-responders. Chronic local inflammation, induced by the N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) carcinogen, led to TLS formation with recruitment and differentiation of the immunosuppressive atypical B-cell (ABC) subset within the bladder microenvironment, predominantly in aging female mice compared to their male counterparts. Depletion of ABCs simultaneous to BCG treatment delayed cancer progression in female mice. Our findings provide evidence indicating a role for ABCs in BCG response and will inform future development of therapies targeting the B cell-exhaustion axis.
RESUMO
Sex and age associated differences in the tumor immune microenvironment of non-muscle invasive bladder (NMIBC) cancer and associated clinical outcomes are emerging indicators of treatment outcomes. The incidence of urothelial carcinoma of the bladder is four times higher in males than females; however, females tend to present with a more aggressive disease, a poorer response to immunotherapy and suffer worse clinical outcomes. Recent findings have demonstrated sex differences in the tumor immune microenvironment of non-muscle invasive and muscle invasive bladder cancer and associated clinical outcomes. However, a significant gap in knowledge remains with respect to the current pre-clinical modeling approaches to more precisely recapitulate these differences towards improved therapeutic design. Given the similarities in mucosal immune physiology between humans and mice, we evaluated the sex and age-related immune alterations in healthy murine bladders. Bulk-RNA sequencing and multiplex immunofluorescence-based spatial immune profiling of healthy murine bladders from male and female mice of age groups spanning young to old showed a highly altered immune landscape that exhibited sex and age associated differences, particularly in the context of B cell mediated responses. Spatial profiling of healthy bladders, using markers specific to macrophages, T cells, B cells, activated dendritic cells, high endothelial venules, myeloid cells and the PD-L1 immune checkpoint showed sex and age associated differences. Bladders from healthy older female mice also showed a higher presence of tertiary lymphoid structures (TLSs) compared to both young female and male equivalents. Spatial immune profiling of N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) carcinogen exposed male and female bladders from young and old mice revealed a similar frequency of TLS formation, sex differences in the bladder immune microenvironment and, age associated differences in latency of tumor induction. These findings support the incorporation of sex and age as factors in pre-clinical modeling of bladder cancer and will potentially advance the field of immunotherapeutic drug development to improve clinical outcomes.
Assuntos
Carcinoma de Células de Transição , Neoplasias da Bexiga Urinária , Envelhecimento , Animais , Butilidroxibutilnitrosamina/efeitos adversos , Carcinógenos , Feminino , Humanos , Masculino , Camundongos , Caracteres Sexuais , Microambiente Tumoral , Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/induzido quimicamente , Neoplasias da Bexiga Urinária/patologiaRESUMO
BACKGROUND: To control the spread of SARS-CoV-2 variants of concern (VOCs), Kingston, Frontenac, and Lennox & Addington (KFL&A) Public Health implemented a more stringent COVID-19 case and contact management (CCM) protocol than what was used across Ontario at the time. We describe epidemiological data and public health measures employed during one of the largest COVID-19 outbreaks in the KFL&A region at the time, caused by the SARS-CoV-2 Alpha (B.1.1.7) VOC, to assess this enhanced protocol. METHODS: We obtained line lists of workers associated with the construction site outbreak, and subsequent cases and contacts from case investigators. Case testing, mutation status, and whole genome sequencing were conducted by Public Health Ontario Laboratories. RESULTS: From 409 high-risk contacts of the outbreak, 109 (27%) developed COVID-19. Three generations of spread were associated with the outbreak, affecting seven public health regions across three provinces. Using an enhanced approach to the CCM, KFL&A Public Health caught 15 cases that could have been missed by standard provincial protocols. CONCLUSIONS: Rapid initial spread within the construction site produced a relatively high attack rate among workers (26%) and their immediate contacts (34%). KFL&A Public Health's implementation of stringent CCM protocols and fast testing turn-around time effectively curbed the spread of the disease in subsequent generations - illustrated by the large reduction in attack rate (34%-14%) and cases (50-10) between the second and third generations. Lessons learned from this analysis may inform guidance on the CCM for future SARS-CoV-2 VOCs as well as other highly transmissible communicable diseases.
HISTORIQUE: Pour contrôler la propagation des variants inquiétants (VOC) du SRAS-CoV-2, la région sociosanitaire de Kingston, Frontenac, Lennox et Addington (KFL&A) a adopté un protocole plus rigoureux de gestion des cas et des contacts (GCC) qui était utilisé partout en Ontario à l'époque. Les auteurs décrivent les données épidémiologiques et les mesures sanitaires employées pendant l'une des plus grosses éclosions de COVID-19 de la région sociosanitaire de KFL&A, causée par le VOC Alpha (B.1.1.7) du SRAS-CoV-2, afin d'évaluer ce protocole amélioré. MÉTHODOLOGIE: Les auteurs ont obtenu les listes des lignes des travailleurs associés à l'éclosion sur le chantier de construction, ainsi que des cas et des contacts subséquents des enquêteurs de cas. Les Laboratoires de Santé publique Ontario ont procédé au dépistage des cas et ont vérifié l'état mutationnel et le séquençage du génome entier. RÉSULTATS: Des 409 contacts à haut risque de l'éclosion, 109 (27%) ont contracté la COVID-19. Trois générations de propagation étaient associées à l'éclosion et touchaient sept régions sociosanitaires réparties dans trois provinces. Au moyen d'une approche améliorée de la GCC, la région sociosanitaire de KFL&A a dépisté 15 cas qui auraient pu être omis par les protocoles provinciaux standards. CONCLUSIONS: Une propagation initiale rapide sur le chantier de construction a produit un taux d'attaque relativement élevé chez les travailleurs (26%) et leurs contacts immédiats (34%). Ladoption de protocoles rigoureux de GCC dans la région sociosanitaire de KFL&A et l'obtention rapide des résultats du dépistage ont enrayé la propagation de la maladie avec efficacité dans les générations suivantes, ce qui est démontré par une forte réduction du taux d'attaque (de 34% à 14%) et de cas (de 50 à 10) entre la deuxiéme génération et la troisiéme. Les leçons tirées de cette analyse pourraient éclairer les conseils sur la GCC des futurs VOC du SRAS-COV-2 et des autres maladies contagieuses hautement transmissibles.