Cardiac c-Kit Biology Revealed by Inducible Transgenesis

RATIONALE:Biological significance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant findings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. OBJECTIVE:The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. METHODS AND RESULTS:In vitro studies confirm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2B-EGFP (enhanced green fluorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specific, inducible, and persistent tracking of c-Kit promoter activation. Tagging efficiency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55% lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. CONCLUSIONS:c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published findings

Cardiac c-Kit Biology Revealed by Inducible Transgenesis

RATIONALE:Biological significance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant findings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. OBJECTIVE:The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. METHODS AND RESULTS:In vitro studies confirm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2B-EGFP (enhanced green fluorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specific, inducible, and persistent tracking of c-Kit promoter activation. Tagging efficiency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55% lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. CONCLUSIONS:c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published findings

Science goals and mission architecture of the Europa Lander mission concept

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hand, K., Phillips, C., Murray, A., Garvin, J., Maize, E., Gibbs, R., Reeves, G., San Martin, A., Tan-Wang, G., Krajewski, J., Hurst, K., Crum, R., Kennedy, B., McElrath, T., Gallon, J., Sabahi, D., Thurman, S., Goldstein, B., Estabrook, P., Lee, S. W., Dooley, J. A., Brinckerhoff, W. B., Edgett, K. S., German, C. R., Hoehler, T. M., Hörst, S. M., Lunine, J. I., Paranicas, C., Nealson, K., Smith, D. E., Templeton, A. S., Russell, M. J., Schmidt, B., Christner, B., Ehlmann, B., Hayes, A., Rhoden, A., Willis, P., Yingst, R. A., Craft, K., Cameron, M. E., Nordheim, T., Pitesky, J., Scully, J., Hofgartner, J., Sell, S. W., Barltrop, K. J., Izraelevitz, J., Brandon, E. J., Seong, J., Jones, J.-P., Pasalic, J., Billings, K. J., Ruiz, J. P., Bugga, R. V., Graham, D., Arenas, L. A., Takeyama, D., Drummond, M., Aghazarian, H., Andersen, A. J., Andersen, K. B., Anderson, E. W., Babuscia, A., Backes, P. G., Bailey, E. S., Balentine, D., Ballard, C. G., Berisford, D. F., Bhandari, P., Blackwood, K., Bolotin, G. S., Bovre, E. A., Bowkett, J., Boykins, K. T., Bramble, M. S., Brice, T. M., Briggs, P., Brinkman, A. P., Brooks, S. M., Buffington, B. B., Burns, B., Cable, M. L., Campagnola, S., Cangahuala, L. A., Carr, G. A., Casani, J. R., Chahat, N. E., Chamberlain-Simon, B. K., Cheng, Y., Chien, S. A., Cook, B. T., Cooper, M., DiNicola, M., Clement, B., Dean, Z., Cullimore, E. A., Curtis, A. G., Croix, J-P. de la, Pasquale, P. Di, Dodd, E. M., Dubord, L. A., Edlund, J. A., Ellyin, R., Emanuel, B., Foster, J. T., Ganino, A. J., Garner, G. J., Gibson, M. T., Gildner, M., Glazebrook, K. J., Greco, M. E., Green, W. M., Hatch, S. J., Hetzel, M. M., Hoey, W. A., Hofmann, A. E., Ionasescu, R., Jain, A., Jasper, J. D., Johannesen, J. R., Johnson, G. K., Jun, I., Katake, A. B., Kim-Castet, S. Y., Kim, D. I., Kim, W., Klonicki, E. F., Kobeissi, B., Kobie, B. D., Kochocki, J., Kokorowski, M., Kosberg, J. A., Kriechbaum, K., Kulkarni, T. P., Lam, R. L., Landau, D. F., Lattimore, M. A., Laubach, S. L., Lawler, C. R., Lim, G., Lin, J. Y., Litwin, T. E., Lo, M. W., Logan, C. A., Maghasoudi, E., Mandrake, L., Marchetti, Y., Marteau, E., Maxwell, K. A., Namee, J. B. Mc, Mcintyre, O., Meacham, M., Melko, J. P., Mueller, J., Muliere, D. A., Mysore, A., Nash, J., Ono, H., Parker, J. M., Perkins, R. C., Petropoulos, A. E., Gaut, A., Gomez, M. Y. Piette, Casillas, R. P., Preudhomme, M., Pyrzak, G., Rapinchuk, J., Ratliff, J. M., Ray, T. L., Roberts, E. T., Roffo, K., Roth, D. C., Russino, J. A., Schmidt, T. M., Schoppers, M. J., Senent, J. S., Serricchio, F., Sheldon, D. J., Shiraishi, L. R., Shirvanian, J., Siegel, K. J., Singh, G., Sirota, A. R., Skulsky, E. D., Stehly, J. S., Strange, N. J., Stevens, S. U., Sunada, E. T., Tepsuporn, S. P., Tosi, L. P. C., Trawny, N., Uchenik, I., Verma, V., Volpe, R. A., Wagner, C. T., Wang, D., Willson, R. G., Wolff, J. L., Wong, A. T., Zimmer, A. K., Sukhatme, K. G., Bago, K. A., Chen, Y., Deardorff, A. M., Kuch, R. S., Lim, C., Syvertson, M. L., Arakaki, G. A., Avila, A., DeBruin, K. J., Frick, A., Harris, J. R., Heverly, M. C., Kawata, J. M., Kim, S.-K., Kipp, D. M., Murphy, J., Smith, M. W., Spaulding, M. D., Thakker, R., Warner, N. Z., Yahnker, C. R., Young, M. E., Magner, T., Adams, D., Bedini, P., Mehr, L., Sheldon, C., Vernon, S., Bailey, V., Briere, M., Butler, M., Davis, A., Ensor, S., Gannon, M., Haapala-Chalk, A., Hartka, T., Holdridge, M., Hong, A., Hunt, J., Iskow, J., Kahler, F., Murray, K., Napolillo, D., Norkus, M., Pfisterer, R., Porter, J., Roth, D., Schwartz, P., Wolfarth, L., Cardiff, E. H., Davis, A., Grob, E. W., Adam, J. R., Betts, E., Norwood, J., Heller, M. M., Voskuilen, T., Sakievich, P., Gray, L., Hansen, D. J., Irick, K. W., Hewson, J. C., Lamb, J., Stacy, S. C., Brotherton, C. M., Tappan, A. S., Benally, D., Thigpen, H., Ortiz, E., Sandoval, D., Ison, A. M., Warren, M., Stromberg, P. G., Thelen, P. M., Blasy, B., Nandy, P., Haddad, A. W., Trujillo, L. B., Wiseley, T. H., Bell, S. A., Teske, N. P., Post, C., Torres-Castro, L., Grosso, C. Wasiolek, M. Science goals and mission architecture of the Europa Lander mission concept. The Planetary Science Journal, 3(1), (2022): 22, https://doi.org/10.3847/psj/ac4493.Europa is a premier target for advancing both planetary science and astrobiology, as well as for opening a new window into the burgeoning field of comparative oceanography. The potentially habitable subsurface ocean of Europa may harbor life, and the globally young and comparatively thin ice shell of Europa may contain biosignatures that are readily accessible to a surface lander. Europa's icy shell also offers the opportunity to study tectonics and geologic cycles across a range of mechanisms and compositions. Here we detail the goals and mission architecture of the Europa Lander mission concept, as developed from 2015 through 2020. The science was developed by the 2016 Europa Lander Science Definition Team (SDT), and the mission architecture was developed by the preproject engineering team, in close collaboration with the SDT. In 2017 and 2018, the mission concept passed its mission concept review and delta-mission concept review, respectively. Since that time, the preproject has been advancing the technologies, and developing the hardware and software, needed to retire risks associated with technology, science, cost, and schedule.K.P.H., C.B.P., E.M., and all authors affiliated with the Jet Propulsion Laboratory carried out this research at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration (grant No. 80NM0018D0004). J.I.L. was the David Baltimore Distinguished Visiting Scientist during the preparation of the SDT report. JPL/Caltech2021