title: MIF/CD74 axis is a target for novel therapies in colon carcinomatosis.
abstract: Strategies aimed at obtaining a complete cytoreduction are needed to improve long-term survival for patients with colorectal cancer peritoneal carcinomatosis (CRC-pc).
  
  
  
  We established organoid models from peritoneal metastases of two naïve CRC patients. A standard paraffin inclusion was conducted to compare their 3D structure and immunohistochemical profile with that of the corresponding surgical samples. RNA expression levels of the CRC stem cell marker LGR5 was measured by in situ hybridization. The secretome of organoids was profiled by mass spectrometry. Energy homeostasis of organoids was interfered with 4-IPP and metformin. Biochemical and metabolic changes after drug treatments were investigated by western blot and mass spectrometry. Mitochondria impairment was evaluated by electron microscopy and mitotraker staining.
  
  
  
  The two organoids recapitulated their corresponding clinical samples in terms of 3D structure and immmunoistochemical profile and were positive for the cancer stem cells marker LGR5. Proteomic analyses of organoids highlighted their strong dependence on energy producing pathways, which suggest that their targeting could be an effective therapeutic approach. To test this hypothesis, we treated organoids with two drugs that target metabolism acting on AMP-activated protein kinase (AMPK), the main regulator of cellular energy homeostasis, which may act as metabolic tumour suppressor in CRC. Organoids were treated with 4-IPP, an inhibitor of MIF/CD74 signalling axis which activates AMPK function, or metformin that inhibits mitochondrial respiratory chain complex I. As a new finding we observed that treatment with 4-IPP downregulated AMPK signalling activity, reduced AKT phosphorylation and activated a JNK-mediated stress-signalling response, thus generating mitochondrial impairment and cell death. Metformin treatment enhanced AMPK activation, decreasing the activity of the anabolic factors ribosomal protein S6 and p4EBP-1 and inducing mitochondrial depolarization.
  
  
  
  We provide evidence that the modulation of AMPK activity may be a strategy for targeting metabolism of CRC-pc organoids.
authors.affiliation: Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Bozzi
authors.affiliation: Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Amadeo 42, Milan, 20133, Italy.|Molecular Genetics of Cancer, Fondazione Istituto FIRC di Oncologia Molecolare, via Adamello 16, Milan, 20139, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Mogavero
authors.affiliation: Proteomics Laboratory Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Amadeo 42, Milan, 20133, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Varinelli
authors.affiliation: Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Antonino
authors.initials: A
authors.lastname: Belfiore
authors.affiliation: Department of Predictive and Preventive Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Amadeo 42, Milan, 20133, Italy.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Manenti
authors.affiliation: Laboratory of Clinical Pathology and Medical Genetics, Fondazione IRCCS 'Carlo Besta' Istituto Neurologico, via G. Amadeo 42, Milan, 20133, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Caccia
authors.affiliation: Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Chiara C
authors.initials: CC
authors.lastname: Volpi
authors.affiliation: Fondazione Istituto FIRC di Oncologia Molecolare, via Adamello 16, Milan, 20139, Italy.
authors.forename: Galina V
authors.initials: GV
authors.lastname: Beznoussenko
authors.affiliation: Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Milione
authors.affiliation: Laboratory of Clinical Pathology and Medical Genetics, Fondazione IRCCS 'Carlo Besta' Istituto Neurologico, via G. Amadeo 42, Milan, 20133, Italy.
authors.forename: Valerio
authors.initials: V
authors.lastname: Leoni
authors.affiliation: Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Annunziata
authors.initials: A
authors.lastname: Gloghini
authors.affiliation: Fondazione Istituto FIRC di Oncologia Molecolare, via Adamello 16, Milan, 20139, Italy.
authors.forename: Alexandre A
authors.initials: AA
authors.lastname: Mironov
authors.affiliation: Colorectal Cancer Unit-Department of Surgery, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Ermanno
authors.initials: E
authors.lastname: Leo
authors.affiliation: Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Venezian 1, Milan, 20133, Italy.
authors.forename: Silvana
authors.initials: S
authors.lastname: Pilotti
authors.affiliation: Molecular Genetics of Cancer, Fondazione Istituto FIRC di Oncologia Molecolare, via Adamello 16, Milan, 20139, Italy.
authors.forename: Marco A
authors.initials: MA
authors.lastname: Pierotti
authors.affiliation: Proteomics Laboratory Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Amadeo 42, Milan, 20133, Italy. italia.bongarzone@istitutotumori.mi.it.
authors.forename: Italia
authors.initials: I
authors.lastname: Bongarzone
authors.affiliation: Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via G. Amadeo 42, Milan, 20133, Italy.|Molecular Genetics of Cancer, Fondazione Istituto FIRC di Oncologia Molecolare, via Adamello 16, Milan, 20139, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Gariboldi
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D004734:Energy Metabolism / Q000187:drug effects; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D008263:Macrophage Migration-Inhibitory Factors / Q000378:metabolism*; D008687:Metformin / Q000494:pharmacology*; D058990:Molecular Targeted Therapy; D010534:Peritoneal Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000556:secondary*; D040901:Proteomics; D011743:Pyrimidines / Q000494:pharmacology*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured
chemical_list: C532251:4-iodo-6-phenylpyrimidine; D000944:Antigens, Differentiation, B-Lymphocyte; D000949:Histocompatibility Antigens Class II; C116884:LGR5 protein, human; D008263:Macrophage Migration-Inhibitory Factors; D011743:Pyrimidines; D043562:Receptors, G-Protein-Coupled; C045084:invariant chain; D008687:Metformin; D055372:AMP-Activated Protein Kinases; D019746:Intramolecular Oxidoreductases; C515598:MIF protein, human
keywords: 4-IPP; AMPK; Macrophage migration inhibitory factor; Metabolism; Metformin; Organoids