A drug screening to identify novel combinatorial strategies for boosting cancer immunotherapy efficacy

Abstract

Background Chimeric antigen receptor (CAR) T cells and immune checkpoint blockades (ICBs) have made remarkable breakthroughs in cancer treatment, but the efficacy is still limited for solid tumors due to tumor antigen heterogeneity and the tumor immune microenvironment. The restrained treatment efficacy prompted us to seek new potential therapeutic methods. Methods In this study, we conducted a small molecule compound library screen in a human BC cell line to identify whether certain drugs contribute to CAR T cell killing. Signaling pathways of tumor cells and T cells affected by the screened drugs were predicted via RNA sequencing. Among them, the antitumor activities of JK184 in combination with CAR T cells or ICBs were evaluated in vitro and in vivo. Results We selected three small molecule drugs from a compound library, among which JK184 directly induces tumor cell apoptosis by inhibiting the Hedgehog signaling pathway, modulates B7-H3 CAR T cells to an effector memory phenotype, and promotes B7-H3 CAR T cells cytokine secretion in vitro. In addition, our data suggested that JK184 exerts antitumor activities and strongly synergizes with B7-H3 CAR T cells or ICBs in vivo. Mechanistically, JK184 enhances B7-H3 CAR T cells infiltrating in xenograft mouse models. Moreover, JK184 combined with ICB markedly reshaped the tumor immune microenvironment by increasing effector T cells infiltration and inflammation cytokine secretion, inhibiting the recruitment of MDSCs and the transition of M2-type macrophages in an immunocompetent mouse model. Conclusion These data show that JK184 may be a potential adjutant in combination with CAR T cells or ICB therapy.

Sensor Array-Enabled Identification of Drugs for Repolarization of Macrophages to Anti-Inflammatory Phenotypes

“…Considering drug tolerance and resistance, it is very important to keep developing and identifying new drugs or repositioning “old” drugs. , Therefore, we used PONI-BA-pyrene to detect the effect of seven commonly used immunotherapy agents on macrophage M1-to-M2 repolarization (Table ). Two reported anti-inflammatory drugs, quercetin and metformin, that can induce M2 repolarization were selected as positive controls. , Quercetin induces M1-to-M2 repolarization through AMP-activated protein kinase (AMPK) and Akt signaling pathways, while metformin induces M2 repolarization through the AMPK/mTOR/NLRP3 inflammasome singling pathway…”

“…Considering drug tolerance and resistance, it is very important to keep developing and identifying new drugs or repositioning "old" drugs. 61,62 Therefore, we used PONI-BA-pyrene to detect the effect of seven commonly used immunotherapy agents on macrophage M1-to-M2 repolarization (Table 2). Two reported anti-inflammatory drugs, quercetin and metformin, that can induce M2 repolarization were selected as positive controls.…”

A high-content screen of FDA approved drugs to enhance CAR T cell function: ingenol-3-angelate improves B7-H3-CAR T cell activity by upregulating B7-H3 on the target cell surface via PKCα activation

“…Other examples of drug plus CAR T cell screens have identified hedgehog pathway inhibitors (e.g. JK184) to enhance B7-H3-CAR T cell killing of breast cancer cells via inhibiting tumor resistance to apoptosis [ 28 ], and insulin-like growth factor receptor 1/insulin receptor inhibitors (BMS-754807, linsitinib) to enhance GD2-CAR T cell activity against diffuse midline glioma [ 29 ]. Overall, there is a growing body of evidence demonstrating that cutting edge technologies can inform rational combinatorial CAR T cell treatment strategies for multiple cancer types.…”

A high-content screen identified ingenol-3-angelate as an enhancer of B7-H3-CAR T cell activity by increasing B7-H3 protein expression on the target cell surface via PKCα activation

“…22 Other examples of drug plus CAR T cell screens have identified hedgehog pathway inhibitors (e.g. JK184) to enhance B7-H3-CAR T cell killing of breast cancer cells via inhibiting tumor resistance to apoptosis, 23 and insulin-like growth factor receptor 1/insulin receptor inhibitors (BMS-754807, linsitinib) to enhance GD2-CAR T cell activity against diffuse midline glioma. 24 Overall, there is a growing body of evidence demonstrating that cutting edge technologies can inform rational combinatorial CAR T cell treatment strategies for multiple cancer types.…”