Browsing by Author "Pulakanti, Kirthi"

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    ARID3A and ARID3B induce stem promoting pathways in ovarian cancer cells
    (Elsevier, 2020-05-15) Dausinas, Paige; Pulakanti, Kirthi; Rao, Sridhar; Cole, Jennifer M.; Dahl, Richard; Cowden Dahl, Karen D.; Biochemistry and Molecular Biology, School of Medicine
    ARID3A and ARID3B are paralogs from the AT-Rich interactive Domain (ARID) family. ARID3A and ARID3B associate to regulate genes in B-cells and cancer. We were the first to demonstrate that ARID3B regulates stem cell genes and promotes the cancer stem cell phenotype. Importantly, different knockout phenotypes in mice and distinct patterns of expression in adult animals suggests that ARID3A and ARID3B may have unique functions. In addition, high levels of ARID3B but not ARID3A induce cell death. Our goal was to express ARID3A, ARID3B, or both genes at a moderate level (as can be observed in cancer) and then identify ARID3 regulated genes. We transduced ovarian cancer cells with ARID3A-GFP, ARID3B-RFP, or both. RNA-sequencing was conducted. ARID3A and ARID3B regulated nearly identical sets of genes. Few genes (<5%) were uniquely regulated by ARID3A or ARID3B. ARID3A/B induced genes involved in cancer and stem cell processes including: Twist, MYCN, MMP2, GLI2, TIMP3, and WNT5B. We found that ARID3A and ARID3B also induced expression of each other, providing evidence of the cooperativity. While ARID3A and ARID3B likely have unique functions in distinct contexts, they are largely capable of regulating the same stem cell genes in cancer cells. This study provides a comprehensive list of genes and pathways regulated by ARID3A and ARID3B in ovarian cancer cells.
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    The miR-23a∼27a∼24-2 microRNA Cluster Promotes Inflammatory Polarization of Macrophages
    (The American Association of Immunologists, 2021) Boucher, Austin; Klopfenstein, Nathan; Hallas, William Morgan; Skibbe, Jennifer; Appert, Andrew; Jang, Seok Hee; Pulakanti, Kirthi; Rao, Sridhar; Cowden Dahl, Karen D.; Dahl, Richard; Microbiology and Immunology, School of Medicine
    Macrophages are critical for regulating inflammatory responses. Environmental signals polarize macrophages to either a pro-inflammatory (M1) state or an anti-inflammatory (M2) state. We observed that the microRNA cluster mirn23a, coding for miRs-23a~27a~24–2, regulates mouse macrophage polarization. Gene expression analysis of mirn23a deficient myeloid progenitors revealed a decrease in Toll like receptor and interferon signaling. Mirn23a−/− bone marrow derived macrophages (BMDMs) have an attenuated response to lipopolysaccharide (LPS) demonstrating an anti-inflammatory phenotype in mature cells. In vitro, mirn23a−/− BMDMs have decreased M1 responses and an enhanced M2 responses. Overexpression of mirn23a has the opposite effect enhancing M1 and inhibiting M2 gene expression. Interestingly expression of mirn23a miRNAs goes down with inflammatory stimulation and up with anti-inflammatory stimulation suggesting that its regulation prevents locking macrophages into polarized states. M2 polarization of tumor associated macrophages (TAMs) correlates with poor outcome for many tumors, so to determine if there was a functional consequence of mirn23a loss modulating immune cell polarization we assayed syngeneic tumor growth in wildtype and mirn23a−/− mice. Consistent with the increased anti-inflammatory/ immunosuppressive phenotype in vitro, mirn23a−/− mice inoculated with syngeneic tumor cells had worse outcomes compared to wildtype mice. Co-injecting tumor cells with mirn23a−/− BMDMs into wildtype mice phenocopied tumor growth in mirn23a−/− mice supporting a critical role for mirn23a miRNAs in macrophage mediated tumor immunity. Our data demonstrates that mirn23a regulates M1/M2 polarization and suggests that manipulation of mirn23a miRNA can be used to direct macrophage polarization to drive a desired immune response.