Engineering high performing microfluidic system for rapid non-invasive isolation and expansion of circulating tumor-reactive lymphocytes

Project Summary: Ovarian cancer poses a significant burden on patients, society, and healthcare system. Despite the success of cytoreductive treatment and chemotherapy as the frontline treatment, many patients still experience metastases and develop cancer recurrence. Unfortunately, the second-line therapy against recurrent/metastasized ovarian cancer remains ill-defined. Tumor infiltrating lymphocytes (TILs) are rare immune cells that migrate from the bloodstream into the tumor to exert protective antitumor effects. These cells can be isolated from the patient’s solid tumor and have been successfully used in cellular immunotherapy. However, production of TILs requires an accessible tumor and involves invasive surgical procedures to isolate these cells followed by a complicated ex vivo expansion process before they are ready to be used for therapeutic purposes. For ovarian cancer, unfortunately the accessible tumor is usually already removed by cytoreductive treatment and thus unavailable when the recurrent/metastasized tumors are detected. Hence, rapid non-invasive means to isolate rare immune cells from alternative natural reservoirs (i.e., blood circulation) are urgently needed to accelerate the development of effective anti-tumor regimens. Microfluidic platforms are powerful tools for the isolation and non-destructive analysis of rare populations of cells based on inexpensive instrumentation and facile fabrication approaches. Recently, we demonstrated the use of our innovative high-performance microfluidic platform technology for unprecedented enrichment of TILs from tumor tissues as well as other circulating tumor-reactive lymphocytes (cTRLs) from peripheral blood. This technology can isolate rare immune cells with high purity and yields while retaining their therapeutic efficacy after expansion. Since frequency of cTRLs in blood is expected to be very low (< 0.005%), we will further develop our microfluidic systems to enable rapid isolation and expansion of rare ovarian cTRLs directly from the blood of syngeneic animal models and human patients to assess their therapeutic potential. In the proposed project we will engineer microfluidic devices to enable rapid isolation of cTRLs directly from the peripheral blood. Through comprehensive characterization of the isolated cTRLs, we will deploy the design- build-test-learn cycle to identify cell-surface markers that can be targeted to improve the enrichment and potency of cTRLs. The new microfluidic system will permit the culturing of the rare cTRLs for ex vivo expansion. We will elucidate the tumor specificity, reactivity, and therapeutic potency of the expanded cTRLs. Based on syngeneic results, we will also isolate cTRLs from clinical specimens, characterize their molecular signatures, and evaluate their anti-tumor efficacy in patient-derived organoid models. The project deliverables will include a first-in-class microfluidic cell sorting and culturing system for rapid isolation and expansion of cTRLs with the potential to impact the American population by making second-line treatment against ovarian cancer more durable and amenable to patients that do not benefit from conventional front-line treatment.