Fig 2: Schematic pathway hypothesis of the biological effects of SPINT2/HAI-2 in haematological neoplasms (Myelodysplastic Syndrome and Acute Myeloid Leukemia, AML). Methylation results in the inhibition of SPINT2/HAI-2 expression in bone marrow mesenchymal stromal cells from myelodysplastic syndromes (MDS) and de novo AML patients, resulting in increased secretion of HGF with consequent increase in cell adhesion and in survival/growth of hematopoietic cells, mainly of the abnormal MDS and de novo AML cells, and contributing with the functional abnormalities of microenvironment niche and with cancer progression. This figure was created using Servier Medical Art tools (http://www.servier.com)

Fig 3: Matriptase is expressed in patient samples and is associated with myeloma cell survival.(A–C) Nanostring mRNA expression analysis of (A) ST14, (B) SPINT1 and (C) SPINT2 was investigated in primary cells from patients with multiple myeloma (MM, n = 25, red) and monoclonal gammopathy of undetermined significance (MGUS, n = 3, dark blue), and in peripheral blood mononuclear cells (PBMCs, n = 2, green). (D, E) Kaplan-Meier analysis with log-rank test for (D) progression-free and (E) overall survival data from CoMMpass IA14 cases stratified into the upper 10th percentile (10% high, n = 75) and non-expressers (TPM<1.0, n = 281). (F) ST14 expression at diagnosis and last relapse in RNA-sequenced longitudinal CD138+ patient samples from CoMMpass IA14. Significance was determined by Wilcoxon signed-rank test. Abbreviation: ns: not significant. (p > 0.05).

Fig 4: In a bioscaffold (3D tridimensional system culture produced by decellularized bovine bone marrow), SPINT2 silencing induces cell growth, HGF and CXCL12 secretion and matrix extracellular production. Bioscaffold was seeded with shControl or shSPINT bone marrow mesenchymal stromal cell and submitted to (A) Histological staining (40×). (B) HGF immunohistochemistry (100×); (C) CXCL12 immunohistochemistry (100×); (D) Harris’ hematoxylin staining (extracellular matriz production) (100×). The samples were quantified using the National Institutes of Health ImageJ program32 and results presented in the graph. Data are the mean ± SEM of different microscopy slide analysis. P values are indicated

Fig 5: HAI-1 and HAI-2 protein expression in matriptase overexpression and knockdown myeloma cell lines.Protein expression of HAI-1 and HAI-2 was determined using Western blotting. (A) INA-6 and U266 matriptase overexpression (Matriptase) and control (Mock) cells, and (B) RPMI-8226 and JJN-3 matriptase knockdown (shMatriptase) and control (shMock) cells. One representative of at least three independent experiments is shown.