Immune contexture in high-risk soft tissue sarcomas (STS): A planned analysis of the ISG-STS-1001 randomized trial.
Authors: Pasquali Sandro , Castelli Chiara , Collini Paola , Barisella Marta , Romagosa Cleofe , Bague Silvia , Coindre Jean Michel , De Tos Paolo , Braglia Luca , Palmerini Emanuela , Quagliuolo Vittorio , Martin Broto Javier , Lopez-Pousa Antonio , Grignani Giovanni , Blay Jean-Yves , Diaz Beveridge Robert , Merlo Domenico F , Stacchiotti Silvia , Casali Paolo Giovanni , Gronchi Alessandro 
Published: June 2021

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Background: The characteristics of immune contexture and its prognostic and predictive value in STS is left to be understood. This planned analysis of the ISG-STS-1001 trial, which compared neoadjuvant anthracycline + ifosfamide (AI) vs a histology-tailored (HT) chemotherapy (ChT), was aimed at characterizing the immune contexture after neoadjuvant ChT and investigating any association with the risk of recurrence. Methods: Patients registered in the ISG-STS-1001 study (ID: NCT01710176) were included if they had tumor tissue available for Tissue MicroArray (TMA), which was performed in the area of the surgical specimen with the highest lymphocyte infiltrate. The following markers were analyzed with IHC and measured quantitatively: CD3, CD8, PD1, GranzymeB, Foxp3, CD20, CD163, and PDL1. The T-Distributed Stochastic Neighboring Entities (t-SNE) analysis was used to account for the co-expression of IHC markers in each tumor. The prognostic value of each marker for disease-free survival (DFS) was assessed. Results: This analysis was conducted in 256 of 435 study patients. AI and HT neoadjuvant ChT did not result in any different distribution of immune contexture. Conversely, differences were observed between ‘complex’ karyotype STS (ck-STS: LMS, MPNST, UPS, MFS, pleomorphic liposarcoma, and pleomorphic rhabdomyosarcoma) and ‘simple’ karyotype STS (sk-STS: MLPS and SS). Ck-STS were enriched in both CD3+ and CD8+ cells compared to sk-STS. These cells displayed an heterogeneous distribution and were dispersed inside the tumor nest, keeping direct contact with sarcoma cells. Ck-STS also displayed an enrichment in Granzyme B+, and CD163+ cells. PDL1+ cells were occasionally identified and were more frequent in ck-STS, suggesting an immune-related expression. Most STS were negative for CD20+ cells, however, when present these cells were highly represented and organized in tertiary lymphoid-like structure. The t-SNE generated plot clustered tumors, the ‘cold’ mainly including sk-STS and the ‘hot’ mainly composed by ck-STS. In the ‘hot’ group, a cluster of tumors displayed an immune infiltrate enriched with a high number of CD3, CD8, GranzymeB, PD-1, and PDL-1+ cells. When the prognostic value of the immune markers was investigated, the presence of CD20+ cells was the only independent prognostic factor for DFS (HR=0.68, 95%CI 0.52-0.91) in a histology-stratified estimate adjusting for tumor size in cm (HR=1.07, 95%CI 1.03-1.12) and patient age (HR=1.0, 95%CI 0.97-1.02). Conclusions: Immune contexture differed across sarcoma histologies after neoadjuvant ChT, rather than across the two study arms, with ck-STS being marked by a rich immune contexture. While a CD20+ infiltrate was found to be an independent prognostic factor for a better outcome, further analyses are in progress on the prognosis of patients with the richest immune contexture. Clinical trial information: NCT01710176.