High-throughput screening of small molecules targeting Mycobacterium tuberculosis in human iPSC macrophages.

New treatments are still necessary to eradicate tuberculosis disease. Macrophages derived from human induced pluripotent stem cells (hiPSC-Macs) offer a physiological niche to identify potential new drugs in the context of Mycobacterium tuberculosis (Mtb) infection. Here, we describe the scale-up of hiPSC-Macs production in 5-stack chambers for high-throughput drug screening against Mtb. A rate of approximately 100 million hiPSC-Macs was generated with optimal quality for a period of up to 12 weeks. Moreover, the infection model was optimized using a luminescence-based Mtb reporter strain. The assay showed enough sensitivity to identify compounds that could target host-pathogen interactions during Mtb infection. We interrogated a library of 200,000 compounds in Mtb-infected hiPSC-Macs with a Z-score above 0.3 in all plates analyzed. After secondary assays, 223 qualified hits were selected for further progression.