Supplementary Data from Fibulin-3 Is Uniquely Upregulated in Malignant Gliomas and Promotes Tumor Cell Motility and Invasion

Supplementary Data from Fibulin-3 Is Uniquely Upregulated in Malignant Gliomas and Promotes Tumor Cell Motility and Invasio

Suppl Tables I and II from Development of a Function-Blocking Antibody Against Fibulin-3 as a Targeted Reagent for Glioblastoma

Supplemental Table I: Antibodies used for Western blotting and immunohistochemistry (IHC); Supplemental Table II: Sequences used for qRT-PCR primers.</p

Figures S1 to S8 from Development of a Function-Blocking Antibody Against Fibulin-3 as a Targeted Reagent for Glioblastoma

Figure S1: Generation of mAb428.2 antibody; Figure S2: Affinity of mAb428.2 for fibulin-3; Figure S3: mAb428.2 binds to cell-surface and interstitial fibulin-3; Figure S4: mAb428.2 does not cause acute toxicity; Figure S5: Locally-delivered mAb428.2 targets fibulin-3 signaling and tumor growth; Figure S6: IV-delivered mAb428.2 inhibits tumor growth; Figure S7: Effects of mAb428.2 against intracranial tumors; Figure S8: Biodistribution of mAb428.2 in naïve and tumor-bearing mice.</p

Legends of Figs S1 to S8 unmarked from Development of a Function-Blocking Antibody Against Fibulin-3 as a Targeted Reagent for Glioblastoma

Legends of Figs S1 to S8, unmarked version</p

DataSheet_2_The extracellular matrix protein fibulin-3/EFEMP1 promotes pleural mesothelioma growth by activation of PI3K/Akt signaling.pdf

Malignant pleural mesothelioma (MPM) is an aggressive tumor with poor prognosis and limited therapeutic options. The extracellular matrix protein fibulin-3/EFEMP1 accumulates in the pleural effusions of MPM patients and has been proposed as a prognostic biomarker of these tumors. However, it is entirely unknown whether fibulin-3 plays a functional role on MPM growth and progression. Here, we demonstrate that fibulin-3 is upregulated in MPM tissue, promotes the malignant behavior of MPM cells, and can be targeted to reduce tumor progression. Overexpression of fibulin-3 increased the viability, clonogenic capacity and invasion of mesothelial cells, whereas fibulin-3 knockdown decreased these phenotypic traits as well as chemoresistance in MPM cells. At the molecular level, fibulin-3 activated PI3K/Akt signaling and increased the expression of a PI3K-dependent gene signature associated with cell adhesion, motility, and invasion. These pro-tumoral effects of fibulin-3 on MPM cells were disrupted by PI3K inhibition as well as by a novel, function-blocking, anti-fibulin-3 chimeric antibody. Anti-fibulin-3 antibody therapy tested in two orthotopic models of MPM inhibited fibulin-3 signaling, resulting in decreased tumor cell proliferation, reduced tumor growth, and extended animal survival. Taken together, these results demonstrate for the first time that fibulin-3 is not only a prognostic factor of MPM but also a relevant molecular target in these tumors. Further development of anti-fibulin-3 approaches are proposed to increase early detection and therapeutic impact against MPM.</p

DataSheet_1_The extracellular matrix protein fibulin-3/EFEMP1 promotes pleural mesothelioma growth by activation of PI3K/Akt signaling.pdf

Malignant pleural mesothelioma (MPM) is an aggressive tumor with poor prognosis and limited therapeutic options. The extracellular matrix protein fibulin-3/EFEMP1 accumulates in the pleural effusions of MPM patients and has been proposed as a prognostic biomarker of these tumors. However, it is entirely unknown whether fibulin-3 plays a functional role on MPM growth and progression. Here, we demonstrate that fibulin-3 is upregulated in MPM tissue, promotes the malignant behavior of MPM cells, and can be targeted to reduce tumor progression. Overexpression of fibulin-3 increased the viability, clonogenic capacity and invasion of mesothelial cells, whereas fibulin-3 knockdown decreased these phenotypic traits as well as chemoresistance in MPM cells. At the molecular level, fibulin-3 activated PI3K/Akt signaling and increased the expression of a PI3K-dependent gene signature associated with cell adhesion, motility, and invasion. These pro-tumoral effects of fibulin-3 on MPM cells were disrupted by PI3K inhibition as well as by a novel, function-blocking, anti-fibulin-3 chimeric antibody. Anti-fibulin-3 antibody therapy tested in two orthotopic models of MPM inhibited fibulin-3 signaling, resulting in decreased tumor cell proliferation, reduced tumor growth, and extended animal survival. Taken together, these results demonstrate for the first time that fibulin-3 is not only a prognostic factor of MPM but also a relevant molecular target in these tumors. Further development of anti-fibulin-3 approaches are proposed to increase early detection and therapeutic impact against MPM.</p

DataSheet_1_Alpha2beta1 Integrin Polymorphism in Diffuse Astrocytoma Patients.pdf

Integrins are heterodimeric transmembrane glycoproteins resulting from the non-covalent association of an α and β chain. The major integrin receptor for collagen/laminin, α2β1 is expressed on a wide variety of cell types and plays an essential role in the adhesion of normal and tumor cells to the extracellular matrix. Integrin-triggered signaling pathways promote the invasion and survival of glioma cells by modifying the brain microenvironment. In this study, we investigated the association of a specific genetic polymorphism of integrin α2β1 with the incidence of diffusely infiltrating astrocytoma and the progression of these tumors. Single-nucleotide polymorphism in intron 7 of the integrin ITGA2 gene was examined in 158 patients and 162 controls using polymerase chain reaction and restriction enzyme analysis. The ITGA2 genotype +/+ (with a BglII restriction site in both alleles) exhibited higher frequency in grade II astrocytoma compared to control (P = 0.02) whereas the genotype -/- (lacking the BglII site) correlated with the poorest survival rate (P = 0.04). In addition, in silico analyses of ITGA2 expression from low-grade gliomas (LGG, n = 515) and glioblastomas (GBM, n = 159) indicated that the higher expression of ITGA2 in LGG was associated with poor overall survival (P < 0.0001). However, the distribution of integrin ITGA2 BglII genotypes (+/+, +/-, -/-) was not significantly different between astrocytoma subgroups III and IV (P = 0.65, 0.24 and 0.33; 0.29, 0.48, 0.25, respectively) compared to control. These results suggest a narrow association between the presence of this SNP and indicate that further studies with larger samples are warranted to analyze the relation between tumor grade and overall survival, highlighting the importance of determining these polymorphisms for prognosis of astrocytomas.</p

Additional file 2 of Global DNA methylation profiling reveals chromosomal instability in IDH-mutant astrocytomas

Additional file 2. Fig. 2 Kaplan-Meier analysis of the full test cohort demonstrating significant differences in progression-free (A) and overall survival (B) between cases stratified by 2021 WHO grad

Additional file 3 of Global DNA methylation profiling reveals chromosomal instability in IDH-mutant astrocytomas

Additional file 3. Table 1 List of IDH-mutant astrocytoma case

Additional file 1 of Global DNA methylation profiling reveals chromosomal instability in IDH-mutant astrocytomas

Additional file 1. Fig. 1 Copy number variation differences between initial biopsies and tumor recurrences in IDH-mutant and IDH-wildtype astrocytomas (A) and IDH-mutant astrocytomas with known CIN and CS status (B