Sep 13, 2022

Volume 17Issue 9p1903-2166
Open Access
Cover In this issue, Sun et al. describe robust differentiation of human pluripotent stem cells to macrophages that can host Mycobacteria abscessus infection, which constitutes an efficient platform for studying pathogen-host reactions and drug discovery. Inspired by immunofluorescent and bright field images in this study, the cover art depicts red-fluorescing-adherent macrophages becoming infected with green GFP-expressing mycobacteria. Cover art by Kathleen Strumila....
Cover In this issue, Sun et al. describe robust differentiation of human pluripotent stem cells to macrophages that can host Mycobacteria abscessus infection, which constitutes an efficient platform for studying pathogen-host reactions and drug discovery. Inspired by immunofluorescent and bright field images in this study, the cover art depicts red-fluorescing-adherent macrophages becoming infected with green GFP-expressing mycobacteria. Cover art by Kathleen Strumila.



  • Regulation of male germline transmission patterns by the Trp53-Cdkn1a pathway

    • Mito Kanatsu-Shinohara,
    • Honda Naoki,
    • Takashi Tanaka,
    • Misako Tatehana,
    • Takako Kikkawa,
    • Noriko Osumi,
    • Takashi Shinohara
    SSCs exhibit an enormous genetic diversity. However, little is known regarding the rules and molecular mechanisms that govern germline transmission patterns. Kanatsu-Shinohara et al. show that TRP53 regulates the male germline transmission pattern by suppressing spermatogonial apoptosis via CDKN1A. Therefore, the TRP53-CDKN1A pathway regulates tumorigenesis and the germline transmission pattern.
  • Exclusive generation of rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells

    • Joel Zvick,
    • Monika Tarnowska-Sengül,
    • Adhideb Ghosh,
    • Nicola Bundschuh,
    • Pjeter Gjonlleshaj,
    • Laura C. Hinte,
    • Christine L. Trautmann,
    • Falko Noé,
    • Xhem Qabrati,
    • Seraina A. Domenig,
    • Inseon Kim,
    • Thomas Hennek,
    • Ferdinand von Meyenn,
    • Ori Bar-Nur
    In this article, Zvick and colleagues used Tsc22d3-deficient mouse embryos to investigate blastocyst complementation of the male germline with mouse and rat pluripotent stem cells. They demonstrate generation of interspecies mouse-rat chimeras exclusively carrying rat PSC-derived germ cells that can fertilize rat oocytes. In addition, they deconstruct xenogeneic rat spermatogenesis occurring in Tsc22d3-deficient mice using scRNA-seq.
  • Rapid tissue prototyping with micro-organospheres

    • Zhaohui Wang,
    • Matteo Boretto,
    • Rosemary Millen,
    • Naveen Natesh,
    • Elena S. Reckzeh,
    • Carolyn Hsu,
    • Marcos Negrete,
    • Haipei Yao,
    • William Quayle,
    • Brook E. Heaton,
    • Alfred T. Harding,
    • Shree Bose,
    • Else Driehuis,
    • Joep Beumer,
    • Grecia O. Rivera,
    • Ravian L. van Ineveld,
    • Donald Gex,
    • Jessica DeVilla,
    • Daisong Wang,
    • Jens Puschhof,
    • Maarten H. Geurts,
    • Athena Yeung,
    • Cait Hamele,
    • Amber Smith,
    • Eric Bankaitis,
    • Kun Xiang,
    • Shengli Ding,
    • Daniel Nelson,
    • Daniel Delubac,
    • Anne Rios,
    • Ralph Abi-Hachem,
    • David Jang,
    • Bradley J. Goldstein,
    • Carolyn Glass,
    • Nicholas S. Heaton,
    • David Hsu,
    • Hans Clevers,
    • Xiling Shen
    Dr. Wang and colleagues have developed micro-organosphere (MOS) technology that establishes miniature 3D tissue models rapidly in a scalable, high-throughput fashion. The unique features of MOSs enable direct viral infection for COVID-19 drug screens, immune cell penetration for a CAR-T potency assay, and assessment of nutrient dependence. Combined with AI-imaging analysis, MOSs offer a clinically feasible assay to guide precision oncology.
  • Whole-genome CRISPR screening identifies genetic manipulations to reduce immune rejection of stem cell-derived islets

    • Elad Sintov,
    • Igor Nikolskiy,
    • Victor Barrera,
    • Jennifer Hyoje-Ryu Kenty,
    • Alexander S. Atkin,
    • Dario Gerace,
    • Shannan J. Ho Sui,
    • Kyle Boulanger,
    • Douglas A. Melton
    Using single-cell transcriptomics and whole-genome CRISPR screening, Sintov and colleagues find gene perturbation targets that can promote immune evasion of SC-islets in allogeneic transplantations. One of these targets, the early inflammatory response chemokine CXCL10, was depleted in SC-islets, which resulted in partial immune evasion in vitro and in vivo.
  • The trophectoderm acts as a niche for the inner cell mass through C/EBPα-regulated IL-6 signaling

    • Marcos Plana-Carmona,
    • Gregoire Stik,
    • Romain Bulteau,
    • Carolina Segura-Morales,
    • Noelia Alcázar,
    • Chris D.R. Wyatt,
    • Antonios Klonizakis,
    • Luisa de Andrés-Aguayo,
    • Maxime Gasnier,
    • Tian V. Tian,
    • Guillem Torcal Garcia,
    • Maria Vila-Casadesús,
    • Nicolas Plachta,
    • Manuel Serrano,
    • Mirko Francesconi,
    • Thomas Graf
    In this article, Graf and colleagues show that an exogenous pulse of the transcription factor C/EBPα activates the IL-6 signaling pathway in B cells, which is required for their efficient reprogramming into iPSCs. This mechanism recapitulates a C/EBPα-IL-6 regulatory axis in the trophectoderm during blastocyst development where the surrounded pluripotent inner cell mass is enriched for the IL-6 receptor.
  • Dual inhibition of MAPK and PI3K/AKT pathways enhances maturation of human iPSC-derived cardiomyocytes

    • Bayardo I. Garay,
    • Sophie Givens,
    • Phablo Abreu,
    • Man Liu,
    • Doğacan Yücel,
    • June Baik,
    • Noah Stanis,
    • Taylor M. Rothermel,
    • Alessandro Magli,
    • Juan E. Abrahante,
    • Natalya A. Goloviznina,
    • Hossam A.N. Soliman,
    • Neha R. Dhoke,
    • Michael Kyba,
    • Patrick W. Alford,
    • Samuel C. Dudley Jr.,
    • Jop H. van Berlo,
    • Brenda Ogle,
    • Rita R.C. Perlingeiro
    In this article, Garay and colleagues show that the MAPK and PI3K/AKT signaling pathways are downregulated in the adult ventricular heart tissue. Inhibition of these signaling pathways in vitro, for only 5 days, can enhance the maturation status of human iPSC-derived cardiomyocytes across multiple domains. This short protocol opens the possibility for synergistic use with other known inducers of cardiac maturation to potentially reach adult-level maturation.
  • Characterization of N-terminal RYR2 variants outside CPVT1 hotspot regions using patient iPSCs reveal pathogenesis and therapeutic potential

    • Marissa J. Stutzman,
    • C.S. John Kim,
    • David J. Tester,
    • Samantha K. Hamrick,
    • Steven M. Dotzler,
    • John R. Giudicessi,
    • Marco C. Miotto,
    • Jeevan B. GC,
    • Joachim Frank,
    • Andrew R. Marks,
    • Michael J. Ackerman
    Ackerman and colleagues provide the first functional evidence that variants N-terminal to the RyR2 mutation hotspot domains alter calcium handling similar to classical CPVT1-causative variants. Using patient-specific induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), they demonstrated that nadolol is effective in reducing spontaneous calcium release associated with RyR2-F13L, -L14P, and -R15P variants.
  • Physiological calcium combined with electrical pacing accelerates maturation of human engineered heart tissue

    • Shi Shen,
    • Lorenzo R. Sewanan,
    • Stephanie Shao,
    • Saiti S. Halder,
    • Paul Stankey,
    • Xia Li,
    • Stuart G. Campbell
    Campbell and colleagues developed a simple method to accelerate maturation of engineered human myocardium that involves culturing tissues under physiological calcium levels while applying a progressively increasing rate of electrical pacing. These tissues display positive force-frequency behavior and improved twitch kinetics, β-adrenergic response, calcium handling, and cardiac troponin I expression within 25 days.
  • P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB

    • Dor Zlotnik,
    • Tatiana Rabinski,
    • Aviv Halfon,
    • Shira Anzi,
    • Inbar Plaschkes,
    • Hadar Benyamini,
    • Yuval Nevo,
    • Orly Yahalom Gershoni,
    • Benyamin Rosental,
    • Eli Hershkovitz,
    • Ayal Ben-Zvi,
    • Gad D. Vatine
    In this article, Vatine and colleagues show that P450 oxidoreductase (POR) is crucial for the development of a functional blood brain barrier (BBB). They show that POR regulates cellular retinoic acid levels through mediating its CYP26-dependent catabolism. In turn, transcriptional pathways involving tight junction and integrin genes are disrupted. These barrier impairments may contribute to the neurological symptoms in POR-deficient patients.
  • The epigenetic state of EED-Gli3-Gli1 regulatory axis controls embryonic cortical neurogenesis

    • Shuang-Feng Zhang,
    • Shang-Kun Dai,
    • Hong-Zhen Du,
    • Hui Wang,
    • Xing-Guo Li,
    • Yi Tang,
    • Chang-Mei Liu
    In this article, Chang-Mei Liu et al. show that the deletion of EED in mouse forebrain leads to disruption of cortical development, decreased proliferation, and increased neuronal differentiation of NSPCs. Furthermore, they identify a novel EED-Gli3-Gli1 regulatory axis that is critical for embryonic brain development, which might lead us to find new therapeutic strategies for neurodevelopmental diseases.
  • SETD4 cells contribute to brain development and maintain adult stem cell reservoir for neurogenesis

    • Sun-Li Cai,
    • Yao-Shun Yang,
    • Yan-Fu Ding,
    • Shu-Hua Yang,
    • Xi-Zheng Jia,
    • Yun-Wen Gu,
    • Chris Wood,
    • Xue-Ting Huang,
    • Jin-Shu Yang,
    • Wei-Jun Yang
    In this article, Yang and colleagues identify a small population of long-lived deep quiescent NSCs in the subventricular zone of the murine brain with lineage tracing of SETD4. These NSCs possess embryonic origins and contribute to brain development before entering quiescence. This quiescent NSC reservoir preserves the capacity of neurogenesis and damage repair once activated.
  • Neuroblast migration along cellular substrates in the developing porcine brain

    • Demisha D.L. Porter,
    • Sara N. Henry,
    • Sadia Ahmed,
    • Amy L. Rizzo,
    • Rita Makhlouf,
    • Collin Gregg,
    • Paul D. Morton
    Porter et al. evaluated the spatiotemporal distribution of immature neurons (neuroblasts) in the subventricular zone of the postnatal piglet. Multiplex labeling revealed that migrating neuroblasts are likely cortical inhibitory interneurons originating from the ventral embryonic forebrain. These findings in the postnatal piglet provide a new platform and viewpoint for understanding neuronal migration and cortical development in normal and diseased states.
  • Loss of endosomal exchanger NHE6 leads to pathological changes in tau in human neurons

    • Marty A. Fernandez,
    • Fatmata Bah,
    • Li Ma,
    • YouJin Lee,
    • Michael Schmidt,
    • Elizabeth Welch,
    • Eric M. Morrow,
    • Tracy L. Young-Pearse
    Fernandez et al. show that loss of the endosomal protein NHE6, mutations in which cause the X-linked neurodevelopmental disorder Christianson syndrome, leads to changes in tau associated with Alzheimer disease and related dementias in iPSC-derived cortical neurons. These neurons display reductions in lysosomal and autophagic function, and the pathological changes in tau are rescued by treatment with trehalose or rapamycin.
  • Human tau mutations in cerebral organoids induce a progressive dyshomeostasis of cholesterol

    • Stella M.K. Glasauer,
    • Susan K. Goderie,
    • Jennifer N. Rauch,
    • Elmer Guzman,
    • Morgane Audouard,
    • Taylor Bertucci,
    • Shona Joy,
    • Emma Rommelfanger,
    • Gabriel Luna,
    • Erica Keane-Rivera,
    • Steven Lotz,
    • Susan Borden,
    • Aaron M. Armando,
    • Oswald Quehenberger,
    • Sally Temple,
    • Kenneth S. Kosik
    In this article, Kosik and colleagues present single-cell RNA sequencing data of cerebral organoids with MAPT mutations causing neurodegeneration. Genes encoding cholesterol biosynthesis enzymes were upregulated in astrocytes of MAPT mutant organoids. Cholesterol and its precursors become elevated as a consequence of MAPT mutations during organoid maturation, suggesting that cholesterol is dysregulated early in disease etiology.


  • Development of a platform to investigate long-term potentiation in human iPSC-derived neuronal networks

    • Deborah Pré,
    • Alexander T. Wooten,
    • Steven Biesmans,
    • Sandy Hinckley,
    • Haowen Zhou,
    • Sean P. Sherman,
    • Priyanka Kakad,
    • Jeffrey Gearhart,
    • Anne G. Bang
    Bang and colleagues describe a multi-electrode array (MEA) assay to investigate LTP on hiPSCderived dopaminergic neuronal networks. Using a chemical LTP paradigm, they demonstrate neuronal network potentiation that lasts for days, partial dependence on BDNF, and induction of canonical activity-regulated gene expression. This platform supports coupling of electrophysiological analyses to studies of LTP molecular mechanisms on hiPSC-derived neuronal networks.
  • Human pluripotent stem cell-derived macrophages host Mycobacterium abscessus infection

    • Shicheng Sun,
    • Michael See,
    • Hieu T. Nim,
    • Kathleen Strumila,
    • Elizabeth S. Ng,
    • Alejandro Hidalgo,
    • Mirana Ramialison,
    • Philip Sutton,
    • Andrew G. Elefanty,
    • Sohinee Sarkar,
    • Edouard G. Stanley
    In this article, Ed Stanley, Sohinee Sarkar, and colleagues develop a simplified method for producing functional macrophages from hPSCs. They use these macrophages to examine intracellular infection with M. abscessus, host cell responses, and antibiotic sensitivity. This model may serve as a system for studying macrophage-pathogen interactions and screening for antimicrobial compounds.