Background The mechanisms by which the intestinal microbiome drives inflammation and disease activity in IBD is not clear. Most of the current efforts in developing treatments targeting the intestinal microbiome in IBD are focused on microbial taxa and very few treatments are developed based on microbiome functions in this condition. Using high-resolution functional microbiome analysis we have designed a live bacterial consortium consisting of four bacterial strains with anti-inflammatory functions (BMC333); and demonstrated the significant anti-inflammatory effects of this consortium in a mouse model of DSS-induced colitis by improving disease activity index, colon length, histologic score and fecal lipocalin (presented at DDW 2022). Aim To validate the anti-inflammatory effects and mechanisms of action of BMC333 using IL10-reporter mouse model. Methods Germ free mice bearing the IL10-GFP reporter gene (B6. IL10-GFP) were orally gavaged with BMC333 at 109 CFU/strain or vehicle/placebo (n=6 in each group) at experimental days 1 and 4. The induction period was followed by 10 days without intervention and the animals were sacrificed at day 14. Immunologic effects was assessed in mouse splenocytes and lamina propria by I. quantitating lamina propria immune-suppressive cells using flow cytometry. II. Comparing the effect of stimulation with each of BMC333 bacterial strains’ lysates on IL10- and IFNγ production by splenocytes from BMC333 or vehicle-treated mice. Results Treatment with BMC333 resulted in: I. Higher levels of immune-suppressive cells (IL10-expressing CD4 T cells and B cells, and regulatory T-cells) in the lamina propria of mice treated with BMC333 compared with vehicle-treated mice. II. Higher numbers of IL10-expressing splenic immunocytes stimulated with lysates compared to non-stimulated cells. Additionally, splenocytes stimulated by each of the BMC333 bacterial strains secreted higher IL10 levels, whereas the strain lysates did not induce secretion of the pro-inflammatory cytokine IFNγ, as measured by ELISA. Conclusion These results validate the anti-inflammatory effects of BMC333 and indicate that this beneficial effect is, at least partially, mediated by increasing IL-10 anti-inflammatory activity as demonstrated by the increased numbers of IL-10-producing lamina propria T and B cells after in vivo colonization, and higher IL-10 producing cells and secretion by BMC333 lysate-stimulated splenocytes. BMC333 stimulates increased immuno-suppressive cell populations in the lamina propria, but no increased production of the pro-inflammatory cytokine IFNγ. These findings are consistent with our previously reported in vivo studies and provide a strong rationale for further development of BMC333 as an effective treatment for IBD.

TPS2685 Background: Immune-Checkpoint inhibitors (ICI) have transformed the treatment of solid tumors, specifically in cutaneous melanoma, clear cell renal cell carcinoma (ccRCC) and non-small cell lung cancer (NSCLC). Despite these advances, a significant proportion of patients do not respond to ICI and only a fraction of responding patients will experience durable responses. Recent reports have found that patients showing poor response to ICI are characterized by a reduced gut microbiome diversity, suggesting that the gut microbiome might affect immune activation by ICI. BMC128 is a live bacterial consortium of 4 bacterial strains predicted to induce anti-tumor immune function when given in conjunction with ICI. These strains were identified by analyzing data obtained from NSCLC and ccRCC patients presenting varied responses to ICI, using a proprietary computational discovery platform enabling high-resolution functional microbiome analysis. In pre-clinical studies, treatment with BMC128 potentiated the efficacy of ICI in breast cancer and melanoma mouse models, reducing tumor volume, increasing the number of responders, and demonstrating an increase in infiltrating immunocytes: CD4, CD8 and NK cells. Methods: In this phase I, first-in-human trial, patients with ccRCC, cutaneous melanoma and NSCLC-adenocarcinoma (EGFR/ALK wild-type), who previously progressed on PD1/PDL-1 inhibitors will be treated with BMC128 8*108 CFU BID in combination with Nivolumab at a fixed-dose of 480mg q4weeks in a 3+3 design, followed by a 9 patient expansion phase. No dose escalation is planned. The primary objective is to investigate the safety and tolerability of BMC128 in combination with Nivolumab. Enrollment is intended to start in Feb, 2022. Clinical trial information: 202122207. [Table: see text]