A High-Throughput Screening Identifies MICU1 Targeting Compounds

  • Cell Rep. 2020 Feb 18;30(7):2321-2331.e6. doi: 10.1016/j.celrep.2020.01.081.
Giulia Di Marco  1 ,  Francesca Vallese  1 ,  Benjamin Jourde  2 ,  Christian Bergsdorf  2 ,  Mattia Sturlese  3 ,  Agnese De Mario  1 ,  Valerie Techer-Etienne  2 ,  Dorothea Haasen  2 ,  Berndt Oberhauser  2 ,  Simone Schleeger  2 ,  Giulia Minetti  2 ,  Stefano Moro  3 ,  Rosario Rizzuto  1 ,  Diego De Stefani  1 ,  Mara Fornaro  4 ,  Cristina Mammucari  5
Affiliations
  • 1. Department of Biomedical Sciences, University of Padua, 35131 Padua, Italy.
  • 2. Novartis Institutes for Biomedical Research, Novartis Campus, 4056 Basel, Switzerland.
  • 3. Molecular Modeling Section, Department of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, Italy.
  • 4. Novartis Institutes for Biomedical Research, Novartis Campus, 4056 Basel, Switzerland. Electronic address: [email protected].
  • 5. Department of Biomedical Sciences, University of Padua, 35131 Padua, Italy. Electronic address: [email protected].
PMID: 32075766 DOI: 10.1016/j.celrep.2020.01.081
Abstract

Mitochondrial CA2+ uptake depends on the mitochondrial calcium uniporter (MCU) complex, a highly selective channel of the inner mitochondrial membrane (IMM). Here, we screen a library of 44,000 non-proprietary compounds for their ability to modulate mitochondrial CA2+ uptake. Two of them, named MCU-i4 and MCU-i11, are confirmed to reliably decrease mitochondrial CA2+ influx. Docking simulations reveal that these molecules directly bind a specific cleft in MICU1, a key element of the MCU complex that controls channel gating. Accordingly, in MICU1-silenced or deleted cells, the inhibitory effect of the two compounds is lost. Moreover, MCU-i4 and MCU-i11 fail to inhibit mitochondrial CA2+ uptake in cells expressing a MICU1 mutated in the critical Amino acids that forge the predicted binding cleft. Finally, these compounds are tested ex vivo, revealing a primary role for mitochondrial CA2+ uptake in muscle growth. Overall, MCU-i4 and MCU-i11 represent leading molecules for the development of MICU1-targeting drugs.

Keywords
HTS; MCU; MICU1; active compounds; high-throughput screening; mitochondrial calcium uniporter; mitochondrial calcium uptake; molecular modeling; small molecules.
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