Courtney Ann Stockman

All Publications

• Efficient Transduction of Alveolar Type 2 Cells with Adeno-associated Virus for the Study of Lung Regeneration AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY Rindler, T. N., Brown, K. M., Stockman, C. A., van Lieshout, L. P., Martin, E. P., Weaver, T. E., Zacharias, W. J., Wootton, S. K., Whitsett, J. A., Bridges, J. P. 2021; 65 (1): 118-121

  View details for Web of Science ID 000672720800016

• Glucocorticoid regulates mesenchymal cell differentiation required for perinatal lung morphogenesis and function AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY Bridges, J. P., Sudha, P., Lipps, D., Wagner, A., Guo, M., Du, Y., Brown, K., Filuta, A., Kitzmiller, J., Stockman, C., Chen, X., Weirauch, M. T., Jobe, A. H., Whitsett, J. A., Xu, Y. 2020; 319 (2): L239-L255

  Abstract

  While antenatal glucocorticoids are widely used to enhance lung function in preterm infants, cellular and molecular mechanisms by which glucocorticoid receptor (GR) signaling influences lung maturation remain poorly understood. Deletion of the glucocorticoid receptor gene (Nr3c1) from fetal pulmonary mesenchymal cells phenocopied defects caused by global Nr3c1 deletion, while lung epithelial- or endothelial-specific Nr3c1 deletion did not impair lung function at birth. We integrated genome-wide gene expression profiling, ATAC-seq, and single cell RNA-seq data in mice in which GR was deleted or activated to identify the cellular and molecular mechanisms by which glucocorticoids control prenatal lung maturation. GR enhanced differentiation of a newly defined proliferative mesenchymal progenitor cell (PMP) into matrix fibroblasts (MFBs), in part by directly activating extracellular matrix-associated target genes, including Fn1, Col16a4, and Eln and by modulating VEGF, JAK-STAT, and WNT signaling. Loss of mesenchymal GR signaling blocked fibroblast progenitor differentiation into mature MFBs, which in turn increased proliferation of SOX9+ alveolar epithelial progenitor cells and inhibited differentiation of mature alveolar type II (AT2) and AT1 cells. GR signaling controls genes required for differentiation of a subset of proliferative mesenchymal progenitors into matrix fibroblasts, in turn, regulating signals controlling AT2/AT1 progenitor cell proliferation and differentiation and identifying cells and processes by which glucocorticoid signaling regulates fetal lung maturation.

  View details for DOI 10.1152/ajplung.00459.2019

  View details for Web of Science ID 000561480000005

  View details for PubMedID 32460513

  View details for PubMedCentralID PMC7473939

• Niche Cells and Signals that Regulate Lung Alveolar Stem Cells In Vivo. Cold Spring Harbor perspectives in biology Juul, N. H., Stockman, C. A., Desai, T. J. 2020

  Abstract

  The distal lung is a honeycomb-like collection of delicate gas exchange sacs called alveoli lined by two interspersed epithelial cell types: the cuboidal, surfactant-producing alveolar type II (AT2) and the flat, gas-exchanging alveolar type I (AT1) cell. During aging, a subset of AT2 cells expressing the canonical Wnt target gene, Axin2, function as stem cells, renewing themselves while generating new AT1 and AT2 cells. Wnt activity endows AT2 cells with proliferative competency, enabling them to respond to activating cues, and simultaneously blocks AT2 to AT1 cell transdifferentiation. Acute alveolar injury rapidly expands the AT2 stem cell pool by transiently inducing Wnt signaling activity in "bulk" AT2 cells, facilitating rapid epithelial repair. AT2 cell "stemness" is thus tightly regulated by access to Wnts, supplied by a specialized single-cell fibroblast niche during maintenance and by AT2 cells themselves during injury repair. Two non-AT2 "reserve" cell populations residing in the distal airways also contribute to alveolar repair, but only after widespread epithelial injury, when they rapidly proliferate, migrate, and differentiate into airway and alveolar lineages. Here, we review alveolar renewal and repair with a focus on the niches, rather than the stem cells, highlighting what is known about the cellular and molecular mechanisms by which they control stem cell activity in vivo.

  View details for DOI 10.1101/cshperspect.a035717

  View details for PubMedID 32179507

• Alveolar injury and regeneration following deletion of ABCA3 JCI INSIGHT Rindler, T. N., Stockman, C. A., Filuta, A. L., Brown, K. M., Snowball, J. M., Zhou, W., Veldhuizen, R., Zink, E. M., Dautel, S. E., Clair, G., Ansong, C., Xu, Y., Bridges, J. P., Whitsett, J. A. 2017; 2 (24)

  Abstract

  Adaptation to air breathing after birth is dependent upon the synthesis and secretion of pulmonary surfactant by alveolar type 2 (AT2) cells. Surfactant, a complex mixture of phospholipids and proteins, is secreted into the alveolus, where it reduces collapsing forces at the air-liquid interface to maintain lung volumes during the ventilatory cycle. ABCA3, an ATP-dependent Walker domain containing transport protein, is required for surfactant synthesis and lung function at birth. Mutations in ABCA3 cause severe surfactant deficiency and respiratory failure in newborn infants. We conditionally deleted the Abca3 gene in AT2 cells in the mature mouse lung. Loss of ABCA3 caused alveolar cell injury and respiratory failure. ABCA3-related lung dysfunction was associated with surfactant deficiency, inflammation, and alveolar-capillary leak. Extensive but incomplete deletion of ABCA3 caused alveolar injury and inflammation, and it initiated proliferation of progenitor cells, restoring ABCA3 expression, lung structure, and function. M2-like macrophages were recruited to sites of AT2 cell proliferation during the regenerative process and were present in lung tissue from patients with severe lung disease caused by mutations in ABCA3. The remarkable and selective regeneration of ABCA3-sufficient AT2 progenitor cells provides plausible approaches for future correction of ABCA3 and other genetic disorders associated with surfactant deficiency and acute interstitial lung disease.

  View details for DOI 10.1172/jci.insight.97381

  View details for Web of Science ID 000418431100015

  View details for PubMedID 29263307

  View details for PubMedCentralID PMC5752264

• A xenograft model of macrophage activation syndrome amenable to anti-CD33 and anti-IL-6R treatment JCI INSIGHT Wunderlich, M., Stockman, C., Devarajan, M., Ravishankar, N., Sexton, C., Kumar, A. R., Mizukawa, B., Mulloy, J. C. 2016; 1 (15): e88181

  Abstract

  Transgenic expression of key myelosupportive human cytokines in immune-deficient mice corrects for the lack of cross-species activities of stem cell factor (SCF), IL-3, and GM-CSF. When engrafted with human umbilical cord blood (UCB), these triple-transgenic mice produce BM and spleen grafts with much higher myeloid composition, relative to nontransgenic controls. Shortly after engraftment with UCB, these mice develop a severe, fatal macrophage activation syndrome (MAS) characterized by a progressive drop in rbc numbers, increased reticulocyte counts, decreased rbc half-life, progressive cytopenias, and evidence of chronic inflammation, including elevated human IL-6. The BM becomes strikingly hypocellular, and spleens are significantly enlarged with evidence of extramedullary hematopoiesis and activated macrophages engaged in hemophagocytosis. This manifestation of MAS does not respond to lymphocyte-suppressive therapies such as steroids, i.v. immunoglobulin, or antibody-mediated ablation of human B and T cells, demonstrating a lymphocyte-independent mechanism of action. In contrast, elimination of human myeloid cells using gemtuzumab ozogamicin (anti-CD33) completely reversed the disease. Additionally, the IL-6R antibody tocilizumab delayed progression and prolonged lifespan. This new model of MAS provides an opportunity for investigation of the mechanisms driving this disease and for the testing of directed therapies in a humanized mouse.

  View details for DOI 10.1172/jci.insight.88181

  View details for Web of Science ID 000387123200010

  View details for PubMedID 27699249

  View details for PubMedCentralID PMC5033750

• Base-modified thymidines capable of terminating DNA synthesis are novel bioactive compounds with activity in cancer cells BIOORGANIC & MEDICINAL CHEMISTRY Borland, K. M., AbdulSalam, S. F., Solivio, M. J., Burke, M. P., Wolfkiel, P. R., Lawson, S. M., Stockman, C. A., Andersen, J. M., Smith, S., Tolstolutskaya, J. N., Gurjar, P. N., Bercz, A. P., Merino, E. J., Litosh, V. A. 2015; 23 (8): 1869-1881

  Abstract

  Current FDA-approved chemotherapeutic antimetabolites elicit severe side effects that warrant their improvement; therefore, we designed compounds with mechanisms of action focusing on inhibiting DNA replication rather than targeting multiple pathways. We previously discovered that 5-(α-substituted-2-nitrobenzyloxy)methyluridine-5'-triphosphates were exquisite DNA synthesis terminators; therefore, we synthesized a library of 35 thymidine analogs and evaluated their activity using an MTT cell viability assay of MCF7 breast cancer cells chosen for their vulnerability to these nucleoside derivatives. Compound 3a, having an α-tert-butyl-2-nitro-4-(phenyl)alkynylbenzyloxy group, showed an IC50 of 9±1μM. The compound is more selective for cancer cells than for fibroblast cells compared with 5-fluorouracil. Treatment of MCF7 cells with 3a elicits the DNA damage response as indicated by phosphorylation of γ-H2A. A primer extension assay of the 5'-triphosphate of 3a revealed that 3aTP is more likely to inhibit DNA polymerase than to lead to termination events upon incorporation into the DNA replication fork.

  View details for DOI 10.1016/j.bmc.2015.01.057

  View details for Web of Science ID 000351852400020

  View details for PubMedID 25778768

  View details for PubMedCentralID PMC4380762