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Purpose of review An update on the latest advances in fertility preservation for transgender women, with an emphasis on the attitudes, access, and techniques. Recent findings With recent warming in perception toward transgendered individuals, fertility preservation services are becoming increasingly available. Although new multidisciplinary transgender clinics are appearing in North America, a 2018 report shows a referral rate for fertility preservation of only 13.5%. Despite interest in child rearing, uptake of fertility preservation is still low as patients are transitioning at increasingly younger ages. Sperm cryopreservation, ideally prior to hormonal therapy, continues to be the mainstay of fertility preservation in transwomen. It is used in conjunction with other assisted reproductive technologies such as intrauterine insemination or in-vitro fertilization with intracytoplasmic sperm injection depending on the quantity of sperm available. Recent research reporting successful autologous grafting of cryopreserved prepubertal testis leading to sperm production and offspring in macaques show great promise in the context of fertility preservation in prepubertal patients. Summary Despite the increasing accessibility of fertility preservation, referral rates and usage continue to be low, suggesting that barriers are still in place for these individuals. Clinicians must continue to counsel patients regarding the process of fertility preservation and advocate for reduction of financial and infrastructural barriers.

High‐grade serous ovarian carcinoma (HGSC) accounts for 70% of all epithelial ovarian cancers but clinical management is challenged by a lack of accurate prognostic and predictive biomarkers of chemotherapy response. This study evaluated the role of Signal Transducer and Activator of Transcription 1 (STAT1) as an independent prognostic and predictive biomarker and its correlation with intratumoural CD8+ T cells in a second independent biomarker validation study. Tumour STAT1 expression and intratumoural CD8+ T cell infiltration were assessed by immunohistochemistry as a multicentre validation study conducted on 734 chemotherapy‐naïve HGSCs. NanoString‐based profiling was performed to correlate expression of STAT1 target genes CXCL9, CXCL10 and CXCL11 with CD8A transcript expression in 143 primary tumours. Multiplexed cytokine analysis of pre‐treatment plasma from resistant and sensitive patients was performed to assess systemic levels of STAT1‐induced cytokines. STAT1 was validated as a prognostic and predictive biomarker in both univariate and multivariate models and its expression correlated significantly with intra‐epithelial CD8+ T cell infiltration in HGSC. STAT1 levels increased the prognostic and predictive value of intratumoural CD8+ T cells, confirming their synergistic role as biomarkers in HGSC. In addition, expression of STAT1 target genes (CXCL9, CXCL10 and CXCL11) correlated significantly with levels of, and CD8A transcripts from intratumoural CD8+ T cells within the resistant and sensitive tumours. Our findings provide compelling evidence that high levels of STAT1, STAT1‐induced chemokines and CD8+ T cells correlate with improved chemotherapy response in HGSC. These results identify STAT1 and its target genes as novel biomarkers of chemosensitivity in HGSC. These findings provide new translational opportunities for patient stratification for immunotherapies based on emerging biomarkers of inflammation in HGSC. An improved understanding of the role of interferon‐inducible genes will be foundational for developing immunomodulatory therapies in ovarian cancer.

Bladder cancer is a management-intensive disease that leads to ~200, 000 deaths worldwide every year. Upon initial diagnosis, approximately 75% of cases are categorized as non-muscle-invasive bladder cancer (NMIBC). Adjuvant Bacillus Calmette-Guérin (BCG) immunotherapy is the gold standard treatment for high risk NMIBC patients. Despite being used for over 40 years, many BCG patients exhibit tumor recurrence, a subset of which display progression to a muscle-invasive form of the disease that requires undesired bladder removal surgery. We recently demonstrated that the interferon response induced by BCG could be further complemented via synergistic activation of the Stimulator of Interferon Genes (STING) pathway. Towards clinical translation of this combinatorial immunomodulation and in order to determine immune biomarkers associated with response to BCG, we retrospectively investigated the immune transcriptomic profiles of tumors from BCG responders and non-responders. Interestingly, tumors from BCG non-responders showed a higher immune cell abundance and higher immune checkpoint gene expression. To further independently validate the transcriptomic findings at protein levels, we conducted spatial immune profiling of 12 immune cell markers (Ki-67+, CD8+ proliferating T cells, FoxP3+ T regulatory cells, CD79+ B cells, PD-1, PD-L1 and IDO1 immune checkpoints, CD68+ and CD163+ M1 and M2 macrophages) in a cohort of 571 NMIBC tumors. Preliminary findings showed that tumors from BCG non-responders are characterized by infiltration of immunosuppressive T regulatory cells. Findings from this study will have the potential to guide future immune biomarker guided immunomodulatory treatment combinations.

Objective We recently established that high STAT1 expression and associated T helper type I tumour immune microenvironment (TME) are prognostic and chemotherapy response predictive biomarkers in high-grade serous ovarian cancer (HGSC). STAT1 induced chemokine CXCL10 is key to the recruitment of lymphocytes in the TME and is significantly highly expressed in the tumours from patients with longer survival. In the current study we therefore aimed to elucidate the role CXCL10 in disease progression and tumour immune transcriptomic alterations using the ID8 syngeneic murine model of HGSC. Methods ID8 ovarian cancer cells were engineered for stable knockdown (KD) and overexpression (OX) of CXCL10. The OX and KD cell line derivatives, along with their respective vector controls, were implanted in immunocompetent C57BL/6 mice via intra-peritoneal injections. At end point, immune transcriptomic profiling of tumour tissues and multiplex cytokine profiling of ascites, was performed. Effect of CXCL10 expression on the tumour vasculature and tumour cell proliferation was evaluated by CD31 and Ki67 immunostaining, respectively. Results Increased CXCL10 expression led to decreased tumour burden and malignant ascites accumulation in the ID8 syngeneic murine model of HGSC. The ascites levels of IL-6 and VEGF were significantly reduced in OX mice compared to the vector controls. The OX tumours also showed reduced vasculature (CD31) and proliferative index (Ki67) compared to the control tumours. Significantly higher expression of genes associated with antigen processing, apoptosis and T cell function was observed in OX tumours compared to the controls. Reduced CXCL10 expression in tumours from KD mice led to increased ascites accumulation and disease progression compared to the controls. Conclusion CXCL10 is a positive determinant of anti-tumour immune responses in HGSC TME and disease progression. These findings are foundational for future translational studies aimed at improving treatment response and survival in HGSC patients, via exploiting the TME.

// Runhan Ren 1, 2 , Kathrin Tyryshkin 3 , Charles H. Graham 1, 2 , Madhuri Koti 2, 4, 5 and D. Robert Siemens 1, 2 1 Department of Urology, Queen’s University, Kingston, ON, Canada 2 Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, ON, Canada 3 Department of Pathology and Molecular Medicine, Queen’s University, Kingston, ON, Canada 4 Cancer Biology and Genetics Division, Queen’s Cancer Research Institute, Queen’s University, Kingston, ON, Canada 5 Department of Obstetrics and Gynecology, Queen’s University, Kingston, ON, Canada Correspondence to: D. Robert Siemens, email: siemensr@kgh.kari.net Keywords: MIBC, immune biomarkers, immunotherapy, TCGA, interferon Received: December 14, 2016 Accepted: May 21, 2017 Published: August 09, 2017 ABSTRACT Recent efforts on genome wide profiling of muscle invasive bladder cancer (MIBC) have led to its classification into distinct genomic and transcriptomic molecular subtypes that exhibit variability in prognosis. Evolving evidence from recent immunotherapy trials has demonstrated the significance of pre-existing tumour immune profiles that could guide treatment decisions. To identify immune gene expression patterns associated with the molecular subtypes, we performed a comprehensive in silico immune transcriptomic profiling, utilizing transcriptomic data from 347 MIBC cases from The Cancer Genome Atlas (TCGA). To investigate subtype-associated immune gene expression patterns, we assembled 924 immune response genes and specifically those involved in T-cell cytotoxicity and the Type I/II interferon pathways. A set of 157 ranked genes was able to distinguish the four subtypes in an unsupervised analysis in an original training cohort (n=122) and an expanded, validation cohort (n=225). The most common overrepresented pathways distinguishing the four molecular subtypes, included JAK/STAT signaling, Toll-like receptor signaling, interleukin signaling, and T-cell activation. Some of the most enriched biological processes were responses to IFN-γ, antigen processing and presentation, cytokine mediated signaling, hemopoeisis, cell proliferation and cellular defense response in the TCGA cluster IV. Our novel findings provide further insights into the association between genomic subtypes and immune activation in MIBC and may open novel opportunities for their exploitation towards precise treatment with immunotherapy.

Background Urothelial bladder cancer (UBC) is a highly prevalent disease in North America, however its optimal management remains elusive. The contribution of B cell associated responses is poorly understood in bladder cancer. Lymphoid neogenesis is a hallmark of an active immune response at tumor sites that sometimes leads to formation of tertiary lymphoid structures (TLS) that resemble germinal centers formed in secondary lymphoid organs. Objective This study was conducted with an aim to investigate the presence and characteristics of TLS in UBC with a focus to compare and contrast the TLS formation in treatment naive low grade non-muscle invasive (NMIBC) and muscle invasive bladder cancers (MIBC). Methods The study cohort consisted of transurethral bladder resection tumour (TURBT) specimens from 28 patients. Sections showing lymphoid aggregates in hematoxylin and eosin (H&E) stained TURBT specimens were further subjected to multi-color immunohistochemistry using immune cell markers specific to CD20+ B cells, CD3+ and CD8+ T cells, PNAd+ high endothelial venules, CD208+ mature dendritic cells, CD21+ follicular dendritic cells to confirm the hallmarks of classical germinal centers. Results Our pilot study investigating the presence of TLS in bladder cancer patients is the first to demonstrate that well-formed TLS are more common in aggressive high grade MIBC tumors compared to low grade NIMBC. Conclusions These novel findings suggest B cell mediated anti-tumour humoral immune responses in bladder cancer progression.

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