Browsing by Author "Chokshi, Mithil"

Now showing 1 - 5 of 5
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    CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages
    (AAAS, 2023) Gillan, Jonathan L.; Chokshi, Mithil; Hardisty, Gareth R.; Clohisey Hendry, Sara; Prasca-Chamorro, Daniel; Robinson, Nicola J.; Lasota, Benjamin; Clark, Richard; Murphy, Lee; Whyte, Moira K. B.; Baillie, J. Kenneth; Davidson, Donald J.; Bao, Gang; Gray, Robert D.
    An intense, nonresolving airway inflammatory response leads to destructive lung disease in cystic fibrosis (CF). Dysregulation of macrophage immune function may be a key facet governing the progression of CF lung disease, but the underlying mechanisms are not fully understood. We used 5′ end centered transcriptome sequencing to profile P. aeruginosa LPS-activated human CF macrophages, showing that CF and non-CF macrophages deploy substantially distinct transcriptional programs at baseline and following activation. This includes a significantly blunted type I IFN signaling response in activated patient cells relative to healthy controls that was reversible upon in vitro treatment with CFTR modulators in patient cells and by CRISPR-Cas9 gene editing to correct the F508del mutation in patient-derived iPSC macrophages. These findings illustrate a previously unidentified immune defect in human CF macrophages that is CFTR dependent and reversible with CFTR modulators, thus providing new avenues in the search for effective anti-inflammatory interventions in CF.
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    Collagen-rich airway smooth muscle cells are a metastatic niche for tumor colonization in the lung
    (Springer Nature, 2019) Lee, Yu-Cheng; Kurtova, Antonina V.; Xiao, Jing; Nikolos, Fotis; Hayashi, Kazukuni; Tramel, Zoe; Jain, Antrix; Chen, Fengju; Chokshi, Mithil; Lee, Ciaran; Bao, Gang; Zhang, Xiang; Shen, Jianjun; Mo, Qianxing; Jung, Sung Yun; Rowley, David; Chan, Keith Syson
    Metastases account for the majority of cancer deaths. While certain steps of the metastatic cascade are well characterized, identification of targets to block this process remains a challenge. Host factors determining metastatic colonization to secondary organs are particularly important for exploration, as those might be shared among different cancer types. Here, we showed that bladder tumor cells expressing the collagen receptor, CD167a, responded to collagen I stimulation at the primary tumor to promote local invasion and utilized the same receptor to preferentially colonize at airway smooth muscle cells (ASMCs)—a rich source of collagen III in lung. Morphologically, COL3-CD167a-driven metastatic foci are uniquely distinct from typical lung alveolar metastatic lesions and exhibited activation of the CD167a-HSP90-Stat3 axis. Importantly, metastatic lung colonization could be abrogated using an investigational drug that attenuates Stat3 activity, implicating this seed-and-soil interaction as a therapeutic target for eliminating lung metastasis.
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    Collagen-rich airway smooth muscle cells are a metastatic niche for tumor colonization in the lung
    (Springer Nature, 2019) Lee, Yu-Cheng; Kurtova, Antonina V.; Xiao, Jing; Nikolos, Fotis; Hayashi, Kazukuni; Tramel, Zoe; Jain, Antrix; Chen, Fengju; Chokshi, Mithil; Lee, Ciaran; Bao, Gang; Zhang, Xiang; Shen, Jianjun; Mo, Qianxing; Jung, Sung Yun; Rowley, David; Chan, Keith Syson
    Metastases account for the majority of cancer deaths. While certain steps of the metastatic cascade are well characterized, identification of targets to block this process remains a challenge. Host factors determining metastatic colonization to secondary organs are particularly important for exploration, as those might be shared among different cancer types. Here, we showed that bladder tumor cells expressing the collagen receptor, CD167a, responded to collagen I stimulation at the primary tumor to promote local invasion and utilized the same receptor to preferentially colonize at airway smooth muscle cells (ASMCs)—a rich source of collagen III in lung. Morphologically, COL3-CD167a-driven metastatic foci are uniquely distinct from typical lung alveolar metastatic lesions and exhibited activation of the CD167a-HSP90-Stat3 axis. Importantly, metastatic lung colonization could be abrogated using an investigational drug that attenuates Stat3 activity, implicating this seed-and-soil interaction as a therapeutic target for eliminating lung metastasis.
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    Genome Editing Strategies to Cure Cystic Fibrosis
    (2022-06-01) Chokshi, Mithil; Bao, Gang
    Cystic Fibrosis (CF) is a recessive genetic disorder with fatal consequences caused by over 1,700 mutations in the Cystic Fibrosis Transmembrane Regulator (CFTR) gene. We studied gene editing in patient-derived induced pluripotent stem cells (iPSCs) by correcting the most prevalent disease-causing mutation, F508del, a 3-bp deletion that prevents CFTR protein to fold properly. The challenges in editing primary and stem cells have caused current approaches to focus on using drug-based selection and fluorescence-based enrichment of the edited cells. We systematically optimized different parameters to obtain highest reported repair rate of 20% allelic HDR at the F508del locus in the iPSCs and conducted an extensive search for off-target activity of the tested gRNA. The cells were then differentiated to epithelia to show recovery of CFTR expression and function. We further optimized the CRISPR / Cas9 machinery to increase the repair rates to above 50%, which is comparable or higher than virus-based editing approaches. To translate our approaches in vivo we are further engineering AAVs to target lung cell types of interest for delivery of the optimized gene editing machinery for an in vivo therapy. These approaches were tested in novel gene-edited human bronchial epithelia-based cell models containing a halide-sensitive fluorescence reporter to conduct high-throughput assays for the CFTR channel activity. We have made such cell models for multiple classes of CF to enable faster discovery of new therapies to cure CF. Finally, CF is a multifaceted disease with fatal lung inflammation and infection cycles; we use gene edited iPSCs developed above to test the hypothesis that CF lung macrophages are hyperinflammatory and poorly bactericidal to perpetuate the inflammation and infection.
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    Human tumor microenvironment chip evaluates the consequences of platelet extravasation and combinatorial antitumor-antiplatelet therapy in ovarian cancer
    (AAAS, 2021) Saha, Biswajit; Mathur, Tanmay; Tronolone, James J.; Chokshi, Mithil; Lokhande, Giriraj K.; Selahi, Amirali; Gaharwar, Akhilesh K.; Afshar-Kharghan, Vahid; Sood, Anil K.; Bao, Gang; Jain, Abhishek
    Platelets extravasate from the circulation into tumor microenvironment, enable metastasis, and confer resistance to chemotherapy in several cancers. Therefore, arresting tumor-platelet cross-talk with effective and atoxic antiplatelet agents in combination with anticancer drugs may serve as an effective cancer treatment strategy. To test this concept, we create an ovarian tumor microenvironment chip (OTME-Chip) that consists of a platelet-perfused tumor microenvironment and which recapitulates platelet extravasation and its consequences. By including gene-edited tumors and RNA sequencing, this organ-on-chip revealed that platelets and tumors interact through glycoprotein VI (GPVI) and tumor galectin-3 under shear. Last, as proof of principle of a clinical trial, we showed that a GPVI inhibitor, Revacept, impairs metastatic potential and improves chemotherapy. Since GPVI is an antithrombotic target that does not impair hemostasis, it represents a safe cancer therapeutic. We propose that OTME-Chip could be deployed to study other vascular and hematological targets in cancer. A tumor microenvironment chip reveals how platelets may fuel cancer metastasis and chemoresistance and unravels a new treatment. A tumor microenvironment chip reveals how platelets may fuel cancer metastasis and chemoresistance and unravels a new treatment.