Multidimensional screening of pancreatic cancer spheroids reveals vulnerabilities in mitotic and cell-matrix adhesion signaling that associate with metastatic progression and decreased patient survival.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy, with a median survival of less than 12 months and a 5-year survival of less than 10 %. Here, we have established an image-based screening pipeline for quantifying single PDAC spheroid dynamics in genetically and phenotypically diverse PDAC cell models. Wild-type KRas PDAC cells formed tight/compact spheroids - compaction of these structures was completely blocked by cytoplasmic dynein and focal adhesion kinase (FAK) inhibitors. In contrast, PDAC cells containing mutant KRas formed loosely aggregated spheroids that grew significantly slower following inhibition of polo-like kinase 1 (PLK1) or focal adhesion kinase (FAK). Independent of genetic background, multicellular PDAC-mesenchymal stromal cell (MSC) spheroids self-organized into structures with an MSC-dominant core. The inclusion of MSCs into wild-type KRas PDAC spheroids modestly affected their compaction; however, MSCs significantly increased the compaction and growth of mutant KRas PDAC spheroids. Notably, exogenous collagen 1 potentiated PANC1 spheroid compaction while ITGA1 knockdown in PANC1 cells blocked MSC-induced PANC1 spheroid compaction. In agreement with a role for collagen-based integrin adhesion complexes in stromal cell-induced PDAC phenotypes, we also discovered that MSC-induced PANC1 spheroid growth was completely blocked by the ITGB1 immunoneutralizing antibody mAb13. Finally, multiplexed single-cell immunohistochemical analysis of a 25 patient PDAC tissue microarray revealed a relationship between decreased variance in Spearman r correlation for ITGA1 and PLK1 expression within the tumor cell compartment of PDAC in patients with advanced disease stage, and elevated expression of both ITGA1 and PLK1 in PDAC was found to be associated with decreased patient survival. Taken together, this work uncovers new therapeutic vulnerabilities in PDAC that are relevant to the progression of this stromal cell-rich malignancy and which may reveal strategies for improving patient outcomes.

The effect of universal leukoreduction on postoperative infections and length of hospital stay in elective orthopedic and cardiac surgery.

BACKGROUND: A before and after study was undertaken to investigate the effect of universal leukoreduction (ULR) in the UK on postoperative length of hospital stay (LOS) and infections. STUDY DESIGN AND METHODS: Consecutive patients undergoing elective coronary artery bypass grafting or total hip and/or knee replacement in 11 hospitals received non-WBC-reduced RBCs before implementation of ULR (T1, n=997) or WBC-reduced RBCs after implementation of ULR (T2, n=1098). RESULTS: Patients in T1 and T2 were comparable except patients in T2 received on average more units of RBCs but had lower discharge Hct levels. Postoperative LOS (T1, 10 +/- 8.9 days; T2, 9.6 +/- 6.9 days) and the proportion of patients with suspected and proven postoperative infections (T1, 21.0%; T2, 20.0%) were unchanged before and after ULR (LOS, hazard ratio 1.01, 95% CI 0.92-1.10; infections, OR 0.83, 95% CI 0.77-1.02). Subgroup analysis showed no significant interaction between storage age or dose of blood on responsiveness of primary outcomes to ULR. Secondary outcomes were unchanged overall. Analysis by surgical procedure gave conflicting results with both increased mortality (p=0.031) and an increased proportion of cardiac patients with proven infections (p=0.004), whereas the proportion of orthopedic patients with proven infections was reduced (p=0.002) after ULR. CONCLUSION: Implementation of ULR had no major impact on postoperative infection or LOS in patients undergoing elective surgical procedures who received transfusion(s). Smaller effects, either detrimental or beneficial of ULR, cannot be excluded.

Endoscopic use of human thrombin in bleeding gastric varices.

OBJECTIVE: The management of gastric variceal hemorrhage remains a clinical challenge. Bovine thrombin has been reported to be effective in two small series. We report our experience with human thrombin in the treatment of bleeding gastric varices. METHODS: We reviewed the case records of 12 patients presenting over a 2-yr period with gastric variceal bleeding requiring endoscopic injection of human thrombin. Ten were male and the mean age was 52 yr (range = 26-83). The underlying diagnoses were cirrhosis in nine, portal vein thrombosis in two, and liver metastasis in one. The majority had fundal gastric varices, and none were thought to have bled from their esophageal varices. Eight received thrombin as primary treatment, whereas four had thrombin only after failing transjugular intrahepatic portosystemic shunts. Patients received one to four sessions (mean = 1.9) of thrombin with a mean total dose of 1833 U (range = 800-4000). Mean follow-up was 17.8 months for those still alive (range = 7-33). RESULTS: Hemostasis in the acute setting was successful in nine patients all of whom received thrombin within 48 hours of the bleed. In the longer term, nine of the 12 had no further bleeding. Of these, five patients did well with thrombin alone, one died of cancer, and the other three went on electively to more definitive shunt procedures. Three patients rebled from their gastric varices of which one was successfully retreated with thrombin. Only one death was related to variceal bleeding (8%). No adverse reactions were noted. CONCLUSION: Our experience demonstrates that endoscopic therapy with thrombin appears safe and can be effective in the management of gastric variceal bleeding.