1. Chemoproteomics and associated bioinformatics.
We have coupled our chemical tools with sensitive mass spectrum-based techniques and bioinformatics to develop a method that provides the cellular complement of proteome alterations associated with a specific stress phenotype. We have conducted several proof-of-principle studies to demonstrate the power of this chemoproteomics approach: identification of an altered signalosome in chronic myeloid leukemia (Nature Chem Biol 2011 and Cell Reports 2015), the viral oncoproteome in AIDS-related lymphomas (Blood 2013), implication of the methyltransferase CARM1 in CML, novel mechanism of activation of STAT5 in cancer (Nature Chem Biol 2011), identification of unanticipated combination therapies for cancer (JCI 2015), novel cancer survival mechanism (Nature 2016), protein networks altered early in the course of Parkinson’s disease (Nature Communications 2018).
Moulick K, Ahn JH, Zong H, Rodina A, Cerchietti L, Gomes DaGama EM, Caldas-Lopes E, Beebe K, Perna F, Hatzi K, Vu LP, Zhao X, Zatorska D, Taldone T, Smith-Jones P, Alpaugh M, Gross SS, Pillarsetty N, Ku T, Lewis JS, Larson SM, Levine R, Erdjument-Bromage H, Guzman ML, Nimer SD, Melnick A, Neckers L, Chiosis G. Affinity-based proteomics reveal cancer-specific networks coordinated by Hsp90. Nat Chem Biol. 2011;7(11):818-26. PubMed PMID: 21946277; PMCID: PMC3265389.
Nature News&Views October 20, 2011, Darby JF, Workman P. Chemical biology: Many faces of a cancer-supporting protein. Nature. 2011;478(7369):334-5. PubMed
2xRecommended F1000Prime: https://f1000.com/prime/13363971
Ojala PM. Naughty chaperone as a target for viral cancer. Blood. 2013;122(16):2767-8. PubMed [PMID: 24136075]
Science News. Tumor-targeting compound points the way to new personalized cancer treatments. December 3, 2011
Adam Bonislawski. ProteoMonitor. Sloan-Kettering Scientists Using Cancer Drug Candidate to Study Aberrant Protein Pathways. Proteomics & Protein Research, Sep 30, 2011
2. Diagnostic assays
These assays are based on our finding that PU-H71 specifically interacts with the tumor HSP90-incorporating epichaperome complexes, such as expressed in certain cancer cells, and other cells under pathogenic stress. We have developed a positron emission tomography based assay that incorporates a radiolabeled version of PU-H71 to detect such epichaperome-positive cells. For liquid tumors we have developed a flow cytometry based assay that incorporates a fluorescently labeled PU-H71, designed for this purpose. The PU-FITC assay for liquid tumors was developed in collaboration with the Guzman lab and the PU-PET assay for solid tumors in collaboration with the Lewis and Larson labs. All assays are currently in clinical evaluation.
3. Compound testing strategies
The distinct nature of the stress chaperome cannot yet be recapitulated biochemically, in a test tube, and we tailor our chemical tools for selectivity towards the CSC through the use of novel, in-house designed and developed, batteries of in silico and in vitro and in vivo phenotypic assays
Rodina A, Taldone T, Kang Y, Patel PD, Koren J, 3rd, Yan P, DaGama Gomes EM, Yang C, Patel MR, Shrestha L, Ochiana SO, Santarossa C, Maharaj R, Gozman A, Cox MB, Erdjument-Bromage H, Hendrickson RC, Cerchietti L, Melnick A, Guzman ML, Chiosis G. Affinity purification probes of potential use to investigate the endogenous Hsp70 interactome in cancer. ACS Chem Biol. 2014;9(8):1698-705. PMCID: PMC4134716.
Corben AD, Uddin MM, Crawford B, Farooq M, Modi S, Gerecitano J, Chiosis G, Alpaugh ML. Ex vivo treatment response of primary tumors and/or associated metastases for preclinical and clinical development of therapeutics. J Vis Exp. 2014(92):e52157. PMCID: PMC4315621.
Taldone T, Patel PD, Patel M, Patel HJ, Evans CE, Rodina A, Ochiana S, Shah SK, Uddin M, Gewirth D, Chiosis G. Experimental and structural testing module to analyze paralogue-specificity and affinity in the Hsp90 inhibitors series. J Med Chem. 2013;56(17):6803-18. PMCID: PMC3985615. PubMed [PMID: 23965125]
Chiosis G, Keeton AB. Assay for isolation of inhibitors of her2-kinase expression. Methods Mol Biol. 2009;486:139-49.
Kang Y, Taldone T, Clement CC, Fewell SW, Aguirre J, Brodsky JL, Chiosis G. Design of a fluorescence polarization assay platform for the study of human Hsp70.Bioorg Med Chem Lett. 2008 May 16.
Chan CT, Paulmurugan R, Gheysens OS, Kim J, Chiosis G, Gambhir SS. Molecular imaging of the efficacy of heat shock protein 90 inhibitors in living subjects. Cancer Res. 2008 Jan 1;68(1):216-26.
Du Y, Moulick K, Rodina A, Aguirre J, Felts S, Dingledine R, Fu H, Chiosis G. High-throughput screening fluorescence polarization assay for tumor-specific Hsp90. J Biomol Screen. 2007 Oct;12(7):915-24.
Moulick K, Clement CC, Aguirre J, Kim J, Kang Y, Felts S, Chiosis G. Synthesis of a red-shifted fluorescence polarization probe for Hsp90. Bioorg Med Chem Lett. 2006 Sep 1;16(17):4515-8. Epub 2006 Jun 22.
Chiosis G, Aguirre J, Nicchitta CV. Synthesis of Hsp90 dimerization modulators. Bioorg Med Chem Lett. 2006 Jul 1;16(13):3529-32. Epub 2006 Apr 18.
Kim J, Felts S, Llauger L, He H, Huezo H, Rosen N, Chiosis G. Development of a fluorescence polarization assay for the molecular chaperone Hsp90. J Biomol Screen. 2004 Aug;9(5):375-81.
Llauger-Bufi L, Felts SJ, Huezo H, Rosen N, Chiosis G.Synthesis of novel fluorescent probes for the molecular chaperone Hsp90. Bioorg Med Chem Lett. 2003 Nov 17;13(22):3975-8.
Huezo H, Vilenchik M, Rosen N, Chiosis G. Microtiter cell-based assay for detection of agents that alter cellular levels of Her2 and EGFR. Chem Biol. 2003 Jul;10(7):629-34.