Cisplatin-based combination chemotherapy (CHT) improves survival in patients with muscle-invasive (MIBC) and metastatic urothelial cancer (mUC). Here we map the cellular landscape in MIBC and mUC and explore its associations to outcome.
We have investigated the amount, proportions and the spatial relationship of tumor and immune cells in MIBC patients receiving neoadjuvant chemotherapy (NAC, n=100) and mUC patients treated with CHT (n=87). We performed multiplex immunofluorescence (mIF) of treatment naive samples on a tissue microarray (TMA), stained with antibodies for CD3, CD163, PD-L1, pSTAT1, CD25, Ki-67, epithelial cells and nuclei. We have developed an image analysis pipeline including machine learning for nuclei cell segmentation and downstream single cells analysis.
Our data using an 8-plex mIF panel confirms that tumor infiltrating lymphocytes is linked to improved prognosis in MIBC. T-cell counts can separate patients into subgroups showing better and worse survival. A higher number of total CD3+ T cells and a higher number of CD3+ T-cells infiltrated into the tumor area associates with better survival in NAC-patients, while this pattern cannot be seen in the mUC patient group. Similarly, a higher number of stroma-infiltrating CD163+ macrophages seems to be a good prognostic factor for the NAC cohort. This is in contrast to the mUC group where a significant worse survival was observed for higher CD163+ cell counts in stroma. We also observed a contrasting survival trend in NAC vs mUC patients with respect to PD-L1-positivity when assessing PDL1-status by a mIF-score mimicking the combined-positive score used for immunohistochemical PD-L1 testing in clinical routine (DAKO 22C3).
mIF enables the exploration of the cellular landscape in advanced urothelial cancer towards clinical endpoints. The spatial distribution of CD3+ and CD163+ cells in tumor and stroma have different prognostic value in mUC and NAC, with possible implications on both treatment and follow-up. Moreover, our results illustrate the dynamic interplay between immune cells and tumor cells during tumor progression.
Anders Ullén and Päivi Östling have equally contributed to this work and share last authorship.
Karolinska Institutet.
Knut och Alice Wallenberg foundation, Swedish Cancer Society, Swedish research council, Cancer society Stockholm, King Gustaf V Jubilee fund.
R.M.P. Turkki: Financial Interests, Personal, Full or part-time Employment: AstraZeneca. L. Paavolainen: Financial Interests, Institutional, Invited Speaker: Orion Pharma. F. Costa Svedman: Non-Financial Interests, Personal, Principal Investigator, Principal investigator för clinical trials in our clinic: MSD; Non-Financial Interests, Personal, Member, SFUO Svensk Förening för Urologisk Onkologi: Swedish Union for Uro-oncology; Non-Financial Interests, Personal, Member, SOF Svensk Oncologisk Förening: Swedish Society of Ocology. O.P. Kallioniemi: Financial Interests, Personal, Invited Speaker: AstraZeneca. A. Ullén: Financial Interests, Personal and Institutional, Funding, Research funding for RWD study on avelumab. Participation in advisory board: Merck. All other authors have declared no conflicts of interest.