title: A pyrazolopyran derivative preferentially inhibits the activity of human cytosolic serine hydroxymethyltransferase and induces cell death in lung cancer cells.
abstract: Serine hydroxymethyltransferase (SHMT) is a central enzyme in the metabolic reprogramming of cancer cells, providing activated one-carbon units in the serine-glycine one-carbon metabolism. Previous studies demonstrated that the cytoplasmic isoform of SHMT (SHMT1) plays a relevant role in lung cancer. SHMT1 is overexpressed in lung cancer patients and NSCLC cell lines. Moreover, SHMT1 is required to maintain DNA integrity. Depletion in lung cancer cell lines causes cell cycle arrest and uracil accumulation and ultimately leads to apoptosis. We found that a pyrazolopyran compound, namely 2.12, preferentially inhibits SHMT1 compared to the mitochondrial counterpart SHMT2. Computational and crystallographic approaches suggest binding at the active site of SHMT1 and a competitive inhibition mechanism. A radio isotopic activity assay shows that inhibition of SHMT by 2.12 also occurs in living cells. Moreover, administration of 2.12 in A549 and H1299 lung cancer cell lines causes apoptosis at LD50 34 μM and rescue experiments underlined selectivity towards SHMT1. These data not only further highlight the relevance of the cytoplasmic isoform SHMT1 in lung cancer but, more importantly, demonstrate that, at least in vitro, it is possible to find selective inhibitors against one specific isoform of SHMT, a key target in metabolic reprogramming of many cancer types.
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Marani
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Paone
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Fiascarelli
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Macone
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Gargano
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Rinaldo
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Giardina
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Valentino
authors.initials: V
authors.lastname: Pontecorvi
authors.affiliation: Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: David
authors.initials: D
authors.lastname: Koes
authors.affiliation: Department of Pharmaceutical Sciences and Drug Discovery Institute, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Lee
authors.initials: L
authors.lastname: McDermott
authors.affiliation: Department of Chemistry and Biochemistry Cristol 63, University of Colorado, Boulder, CO 80302, USA.
authors.forename: Tianyi
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Paiardini
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Contestabile
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, Rome 00185, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cutruzzolà
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D020134:Catalytic Domain; D049109:Cell Proliferation / Q000187:drug effects; D003593:Cytoplasm / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D012696:Glycine Hydroxymethyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation; D011714:Pyrans / Q000737:chemistry*; D011720:Pyrazoles / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D011714:Pyrans; D011720:Pyrazoles; D012333:RNA, Messenger; C031280:pyrazole; D012696:Glycine Hydroxymethyltransferase; C520541:SHMT protein, human
keywords: apoptosis; inhibition; lung cancer; pyrazolopyrans; serine hydroxymethyltransferase