Evaluation of Twist 1 and Aurora-A antibodies via immunohistochemistry in Pancreatic Adenocarcinoma

Nearly 55,000 deaths occur as a result of pancreatic cancer annually, making it the fourth most deadly cancer to date. Patients are often not diagnosed until resentation at a stage IV, contributing to the high death toll. In this study, a Tissue Micro Array (TMA) was stained and analyzed for 2 biomarkers. The TMA contained 27 cases had 3 cores per case; 2 cores were adenocarcinomas of the pancreas and a core with non-cancerous adjacent area of tissue. The TMA was immunostained with antibodies TWIST-1 and AURORA kinase to show potential use as biomarkers to help determine treatment. Immunostaining was done by the Dako-flex system platform. Both positive and negative controls were used to verify staining specificity of the two antibodies. The Aperio Whole slide digital imaging system was used to create a digital image of the TMA, and then computer-assisted morphometric analysis was used to analyze the digital images. Immunostaining of Twist-1 was found in the ductal epithelial carcinoma cells in the pancreas, and also observed in the stroma of ductal adenocarcinoma cases. Stroma in higher stages, stages III and IV, showed higher expression of Twist-1 relative to stages I and II. Conversely, AURORA showed no staining in the stroma and was localized in the nucleus of a few tumor cells undergoing proliferation. TWIST-1 is associated with Epithelial to Mesenchymal transformation (EMT) and the data indicates Twist-1 may play a role in chemo-resistance of pancreatic cancer

Inhibition of BTK and PI3Kδ impairs the development of human JMML stem and progenitor cells

Juvenile myelomonocytic leukemia (JMML) is an aggressive myeloproliferative neoplasia that lacks effective targeted chemotherapies. Clinically, JMML manifests as monocytic leukocytosis, splenomegaly with consequential thrombocytopenia. Most commonly, patients have gain-of-function (GOF) oncogenic mutations in PTPN11 (SHP2), leading to Erk and Akt hyperactivation. Mechanism(s) involved in co-regulation of Erk and Akt in the context of GOF SHP2 are poorly understood. Here, we show that Bruton's tyrosine kinase (BTK) is hyperphosphorylated in GOF Shp2-bearing cells and utilizes B cell adaptor for PI3K to cooperate with p110δ, the catalytic subunit of PI3K. Dual inhibition of BTK and p110δ reduces the activation of both Erk and Akt. In vivo, individual targeting of BTK or p110δ in a mouse model of human JMML equally reduces monocytosis and splenomegaly; however, the combined treatment results in a more robust inhibition and uniquely rescues anemia and thrombocytopenia. RNA-seq analysis of drug-treated mice showed a profound reduction in the expression of genes associated with leukemic cell migration and inflammation, leading to correction in the infiltration of leukemic cells in the lung, liver, and spleen. Remarkably, in a patient derived xenograft model of JMML, leukemia-initiating stem and progenitor cells were potently inhibited in response to the dual drug treatment

Pharmacological inhibition of Carbonic Anhydrase IX and XII to enhance targeting of acute myeloid leukaemia cells under hypoxic conditions

Acute myeloid leukaemia (AML) is an aggressive form of blood cancer that carries a dismal prognosis. Several studies suggest that the poor outcome is due to a small fraction of leukaemic cells that elude treatment and survive in specialised, oxygen (O2)‐deprived niches of the bone marrow. Although several AML drug targets such as FLT3, IDH1/2 and CD33 have been established in recent years, survival rates remain unsatisfactory, which indicates that other, yet unrecognized, mechanisms influence the ability of AML cells to escape cell death and to proliferate in hypoxic environments. Our data illustrates that Carbonic Anhydrases IX and XII (CA IX/XII) are critical for leukaemic cell survival in the O2‐deprived milieu. CA IX and XII function as transmembrane proteins that mediate intracellular pH under low O2 conditions. Because maintaining a neutral pH represents a key survival mechanism for tumour cells in O2‐deprived settings, we sought to elucidate the role of dual CA IX/XII inhibition as a novel strategy to eliminate AML cells under hypoxic conditions. Our findings demonstrate that the dual CA IX/XII inhibitor FC531 may prove to be of value as an adjunct to chemotherapy for the treatment of AML.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/171236/1/jcmm17027_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/171236/2/jcmm17027-sup-0001-FigS1-S4.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/171236/3/jcmm17027.pd