Late diagnosis contributes to pancreatic cancer (PaCa) dismal prognosis, urging for reliable, early detection. Serum‐exosome protein and/or miRNA markers might be suitable candidates, which we controlled for patients with PaCa. Protein markers were selected according to expression in exosomes of PaCa cell line culture supernatants, but not healthy donors' serum‐exosomes. miRNA was selected according to abundant recovery in microarrays of patients with PaCa, but not healthy donors' serum‐exosomes and exosome‐depleted serum. According to these preselections, serum‐exosomes were tested by flow cytometry for the PaCa‐initiating cell (PaCIC) markers CD44v6, Tspan8, EpCAM, MET and CD104. Serum‐exosomes and exosome‐depleted serum was tested for miR‐1246, miR‐4644, miR‐3976 and miR‐4306 recovery by qRT‐PCR. The majority (95%) of patients with PaCa (131) and patients with nonPa‐malignancies reacted with a panel of anti‐CD44v6, ‐Tspan8, ‐EpCAM and ‐CD104. Serum‐exosomes of healthy donors' and patients with nonmalignant diseases were not reactive. Recovery was tumor grading and staging independent including early stages. The selected miR‐1246, miR‐4644, miR‐3976 and miR‐4306 were significantly upregulated in 83% of PaCa serum‐exosomes, but rarely in control groups. These miRNA were also elevated in exosome‐depleted serum of patients with PaCa, but at a low level. Concomitant evaluation of PaCIC and miRNA serum‐exosome marker panels significantly improved sensitivity (1.00, CI: 0.95–1) with a specificity of 0.80 (CI: 0.67–0.90) for PaCa versus all others groups and of 0.93 (CI: 0.81–0.98) excluding nonPa‐malignancies. Thus, the concomitant evaluation of PaCIC and PaCa‐related miRNA marker panels awaits retrospective analyses of larger cohorts, as it should allow for a highly sensitive, minimally‐invasive PaCa diagnostics.

Legumain is required for maintenance of normal kidney homeostasis. However, its role in acute kidney injury (AKI) is still unclear. Here, we induced AKI by bilateral ischemia-reperfusion injury (IRI) of renal arteries or folic acid in lgmn^WT and lgmn^KO mice. We assessed serum creatinine, blood urea nitrogen, histological indexes of tubular injury, and expression of KIM-1 and NGAL. Inflammatory infiltration was evaluated by immunohistological staining of CD3 and F4/80, and expression of TNF-α, CCL-2, IL-33, and IL-1α. Ferroptosis was evaluated by Acsl4, Cox-2, reactive oxygen species (ROS) indexes H₂DCFDA and DHE, MDA and glutathione peroxidase 4 (GPX4). We induced ferroptosis by hypoxia or erastin in primary mouse renal tubular epithelial cells (mRTECs). Cellular survival, Acsl4, Cox-2, LDH release, ROS, and MDA levels were measured. We analyzed the degradation of GPX4 through inhibition of proteasomes or autophagy. Lysosomal GPX4 was assessed to determine GPX4 degradation pathway. Immunoprecipitation (IP) was used to determine the interactions between legumain, GPX4, HSC70, and HSP90. For tentative treatment, RR-11a was administrated intraperitoneally to a mouse model of IRI-induced AKI. Our results showed that legumain deficiency attenuated acute tubular injury, inflammation, and ferroptosis in either IRI or folic acid-induced AKI model. Ferroptosis induced by hypoxia or erastin was dampened in lgmn^KO mRTECs compared with lgmn^WT control. Deficiency of legumain prevented chaperone-mediated autophagy of GPX4. Results of IP suggested interactions between legumain, HSC70, HSP90, and GPX4. Administration of RR-11a ameliorated ferroptosis and renal injury in the AKI model. Together, our data indicate that legumain promotes chaperone-mediated autophagy of GPX4 therefore facilitates tubular ferroptosis in AKI.

Tspan8 and CD151 are metastasis-promoting tetraspanins and a knockdown (kd) of Tspan8 or CD151 and most pronounced of both tetraspanins affects the metastatic potential of the rat pancreatic adenocarcinoma line ASML. Approaching to elaborate the underlying mechanism, we compared ASMLwt, -CD151kd and/or Tspan8kd clones. We focused on tumor exosomes, as exosomes play a major role in tumor progression and tetraspanins are suggested to be engaged in exosome targeting.

Rationale : Polyploidy and multinucleation are characteristic features of mammalian cardiomyocytes, which develop shortly after birth when most differentiated cardiomyocytes become acytokinetic. Cardiac overload and hypertrophy further increase the degree of polyploidy of cardiomyocytes, suggesting a role in cell type–specific responses to physiological and pathological stimuli. Objective : We sought to study the function of cyclinG1 in the regulation of polyploidy and multinucleation in cardiomyocytes. Methods and Results : We found that expression of cyclinG1, a transcriptional target of p53, coincides with arrest of cardiomyocyte proliferation and onset of polyploidization. Overexpression of cyclinG1 promoted DNA synthesis but inhibited cytokinesis in neonatal cardiomyocytes leading to an enlarged population of binuclear cardiomyocytes. Reciprocally, inactivation of the cyclinG1 gene in mice lowered the degree of polyploidy and multinucleation in cardiomyocytes. Moreover, lack of cyclinG1 prevented the increase of polynucleated cardiomyocytes in response to pressure overload and hypertrophy. Conclusions : CyclinG1 is an important player for the regulation of polyploidy and multinucleation in cardiomyocytes probably by inhibition of apoptosis caused by checkpoint activation.

Exosomal tetraspanins mediate cancer metastasis by altering host microenvironment Exosomal tetraspanins mediate cancer metastasis by altering host microenvironment