Stromal androgen and hedgehog signaling regulates stem cell niches in pubertal prostate development

Development ◽  
2021 ◽  
Author(s):  
Adam W. Olson ◽  
Vien Le ◽  
Jinhui Wang ◽  
Alex Hiroto ◽  
Won Kyung Kim ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Growth And Development ◽  
Hedgehog Signaling ◽  
Prostate Development ◽  
Cell Niche ◽  
Cell Trajectory ◽  
Signaling Activation ◽  
Epithelial Growth

Stromal androgen-receptor (AR) action is essential for prostate development, morphogenesis, and regeneration. However, mechanisms underlying how stromal AR maintains the cell niche in support of pubertal prostatic epithelial growth are unknown. Here, using advanced mouse genetic tools, we demonstrate that selective deletion of stromal AR expression in prepubescent Shh responsive Gli1-expressing cells significantly impedes pubertal prostate epithelial growth and development. Single-cell transcriptomic analyses showed that AR loss in these prepubescent Gli1-expressing cells dysregulates androgen-signaling initiated stromal-epithelial paracrine interactions, leading to growth retardation of pubertal prostate epithelia and significant development defects. Specifically, AR loss elevates Shh-signaling activation in both prostatic stromal and adjacent epithelial cells, directly inhibiting prostatic epithelial growth. Single-cell trajectory analyses further identified aberrant differentiation fates of prostatic epithelial cells directly altered by stromal AR deletion. In vivo recombination of AR-deficient stromal Gli1-lineage cells with wild-type prostatic epithelial cells failed to develop normal prostatic epithelia. These data demonstrate novel mechanisms underlying how stromal AR-signaling facilitates Shh-mediated cell niches in pubertal prostatic epithelial growth and development.

scholarly journals Combined transient ablation and single cell RNA sequencing reveals the development of medullary thymic epithelial cells

2020 ◽  
Author(s):  
Kristen L. Wells ◽  
Corey N. Miller ◽  
Andreas R. Gschwind ◽  
Wu Wei ◽  
Jonah D. Phipps ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Expression Patterns ◽  
Critical Role ◽  
Lineage Tracing ◽  
Thymic Epithelial Cells ◽  
Single Cell Rna Sequencing ◽  
Medullary Thymic Epithelial Cells

AbstractMedullary thymic epithelial cells (mTECs) play a critical role in central immune tolerance by mediating negative selection of autoreactive T cells through the collective expression of the peripheral self-antigen compartment, including tissue-specific antigens (TSAs). Recent work has shown that gene expression patterns within the mTEC compartment are remarkably heterogenous and include multiple differentiated cell states. To further define mTEC development and medullary epithelial lineage relationships, we combined lineage tracing and recovery from transient in vivo mTEC ablation with single cell RNA-sequencing. The combination of bioinformatic and experimental approaches revealed a non-stem transit-amplifying population of cycling mTECs that preceded Aire expression. Based on our findings, we propose a branching model of mTEC development wherein a heterogeneous pool of transit-amplifying cells gives rise to Aire- and Ccl21a-expressing mTEC subsets. We further use experimental techniques to show that within the Aire-expressing developmental branch, TSA expression peaked as Aire expression decreased, implying Aire expression must be established before TSA expression can occur. Collectively, these data provide a higher order roadmap of mTEC development and demonstrate the power of combinatorial approaches leveraging both in vivo models and high-dimensional datasets.

scholarly journals In vitro and in vivo development of the human intestinal niche at single cell resolution

2020 ◽  
Author(s):  
Michael Czerwinski ◽  
Emily M. Holloway ◽  
Yu-Hwai Tsai ◽  
Angeline Wu ◽  
Qianhui Yu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Single Cell ◽  
Gene Signature ◽  
Muscularis Mucosa ◽  
Erbb Signaling ◽  
Forming Efficiency ◽  
Cell Niche ◽  
Niche Factor

SUMMARYThe human intestinal stem cell (ISC) niche supports ISC self-renewal and epithelial function, yet little is known about the development of the human ISC niche. We used single-cell mRNA sequencing (scRNA-seq) to interrogate the human intestine across 7-21 weeks of gestation. Using these data coupled with marker validation in situ, molecular identities and spatial locations were assigned to several cell populations that comprise the epithelial niche, and the cellular origins of many niche factors were determined. The major source of WNT and RSPONDIN ligands were ACTA2+ cells of the muscularis mucosa. EGF was predominantly expressed in the villus epithelium and the EGF-family member NEUREGULIN1 (NRG1) was expressed by subepithelial mesenchymal cells. Functional data from enteroid cultures showed that NRG1 improved cellular diversity, enhanced the stem cell gene signature, and increased enteroid forming efficiency, whereas EGF supported a secretory gene expression profile and stimulated rapid proliferation. This work highlights unappreciated complexities of intestinal EGF/ERBB signaling and identifies NRG1 as a stem cell niche factor.

scholarly journals Set7 mediated Gli3 methylation plays a positive role in the activation of Sonic Hedgehog pathway in mammals

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Lin Fu ◽  
Hailong Wu ◽  
Steven Y Cheng ◽  
Daming Gao ◽  
Lei Zhang ◽  
...  
Keyword(s):  
Hedgehog Signaling ◽  
Positive Role ◽  
Shh Signaling ◽  
Cubitus Interruptus ◽  
Tumor Growth And Metastasis ◽  
Double Agent ◽  
The Stability ◽  
Signaling Activation

Hedgehog signaling plays very important roles in development and cancers. Vertebrates have three transcriptional factors, Gli1, Gli2 and Gli3. Among them, Gli3 is a very special transcriptional factor which closely resembles Cubitus interruptus (Ci, in Drosophila) structurally and functionally as a ‘double agent’ for Shh target gene expression. Here we show that Gli3 full-length, but not the truncated form, can be methylated at K436 and K595. This methylation is specifically catalyzed by Set7, a lysine methyltransferase (KMT). Methylation at K436 and K595 respectively increases the stability and DNA binding ability of Gli3, resulting in an enhancement of Shh signaling activation. Furthermore, functional experiments indicate that the Gli3 methylation contributes to the tumor growth and metastasis in non-small cell lung cancer in vitro and in vivo. Therefore, we propose that Set7 mediated methylation is a novel PTM of Gli3, which positively regulates the transactivity of Gli3 and the activation of Shh signaling.

scholarly journals Wnt Secretion from Epithelial Cells and Subepithelial Myofibroblasts Is Not Required in the Mouse Intestinal Stem Cell Niche In Vivo

2014 ◽  
Vol 2 (2) ◽  
pp. 127-134 ◽  
Author(s):  
Adrianna K. San Roman ◽  
Chenura D. Jayewickreme ◽  
L. Charles Murtaugh ◽  
Ramesh A. Shivdasani
Keyword(s):  
Stem Cell ◽  
Epithelial Cells ◽  
Stem Cell Niche ◽  
Intestinal Stem Cell ◽  
Cell Niche

scholarly journals Disruption of Prostate Epithelial Androgen Receptor Impedes Prostate Lobe-Specific Growth and Function

Endocrinology ◽  
2007 ◽  
Vol 148 (5) ◽  
pp. 2264-2272 ◽  
Author(s):  
Ulla Simanainen ◽  
Charles M. Allan ◽  
Patrick Lim ◽  
Stephen McPherson ◽  
Mark Jimenez ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Epithelial Cells ◽  
Molecular Mechanisms ◽  
Functional Differentiation ◽  
Branching Morphogenesis ◽  
Testis Weight ◽  
Structural Differentiation ◽  
Prostate Development ◽  
Prostate Epithelium

Prostate development and maturation requires stromal-epithelial interactions and androgen action via the androgen receptor (AR) within these compartments. However, the specific roles of epithelial and stromal AR in postnatal prostate differentiation are unclear. We used Cre-LoxP technology to determine the prostate phenotype in mice with epithelial-selective genetic inactivation of the AR leaving the stromal AR functionally intact. We find that prostate development abolished in mice globally lacking a functional AR can be rescued by restricting the AR knockout to the postnatal prostate epithelium. We show that, at 8 wk of age, prostate epithelial AR knockout (PEARKO) mice exhibit prostate development with normal branching morphogenesis but lobe-specific decrease in prostate weight and hindered structural and functional differentiation of the mature prostate epithelium. No change was observed in PEARKO testis weight or serum testosterone compared with littermate controls. The most striking change was increased proliferation and abnormal lesions of epithelial cells predominantly in the anterior lobe of PEARKO mice. These findings highlight the vital role of stromal AR in postnatal prostate growth and structural differentiation and emphasize the requirement of epithelial AR in maintaining functional differentiation and restraining proliferation of epithelial cells in a lobe-specific manner. This unique PEARKO mouse provides a new paradigm with which to define the molecular mechanisms of the androgen signaling in mature prostate lobes in vivo and provides insight into the identification of better targets for treatment of prostate cancer and hyperplasia.

scholarly journals A novel protein encoded by circular SMO RNA is essential for Hedgehog signaling activation and glioblastoma tumorigenicity

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Xujia Wu ◽  
Songhua Xiao ◽  
Maolei Zhang ◽  
Lixuan Yang ◽  
Jian Zhong ◽  
...  
Keyword(s):  
Hedgehog Signaling ◽  
Transmembrane Receptors ◽  
Novel Target ◽  
Signaling Activation ◽  
Cholesterol Modification ◽  
G Protein Coupled ◽  
Proliferation In Vitro ◽  
Novel Protein

Abstract Background Aberrant activation of the Hedgehog pathway drives tumorigenesis of many cancers, including glioblastoma. However, the sensitization mechanism of the G protein-coupled-like receptor smoothened (SMO), a key component of Hedgehog signaling, remains largely unknown. Results In this study, we describe a novel protein SMO-193a.a. that is essential for Hedgehog signaling activation in glioblastoma. Encoded by circular SMO (circ-SMO), SMO-193a.a. is required for sonic hedgehog (Shh) induced SMO activation, via interacting with SMO, enhancing SMO cholesterol modification, and releasing SMO from the inhibition of patched transmembrane receptors. Deprivation of SMO-193a.a. in brain cancer stem cells attenuates Hedgehog signaling intensity and suppresses self-renewal, proliferation in vitro, and tumorigenicity in vivo. Moreover, circ-SMO/SMO-193a.a. is positively regulated by FUS, a direct transcriptional target of Gli1. Shh/Gli1/FUS/SMO-193a.a. form a positive feedback loop to sustain Hedgehog signaling activation in glioblastoma. Clinically, SMO-193a.a. is more specifically expressed in glioblastoma than SMO and is relevant to Gli1 expression. Higher expression of SMO-193a.a. predicts worse overall survival of glioblastoma patients, indicating its prognostic value. Conclusions Our study reveals that SMO-193a.a., a novel protein encoded by circular SMO, is critical for Hedgehog signaling, drives glioblastoma tumorigenesis and is a novel target for glioblastoma treatment.

scholarly journals Single-cell profiling and SCOPE-seq reveal the lineage dynamics of adult neurogenesis and NOTUM as a key V-SVZ regulator

2019 ◽  
Author(s):  
Dogukan Mizrak ◽  
N. Sumru Bayin ◽  
Jinzhou Yuan ◽  
Zhouzerui Liu ◽  
Radu Suciu ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Single Cell ◽  
Temporal Dynamics ◽  
Dynamic Control ◽  
Mammalian Brain ◽  
Regulatory State ◽  
Surface Binding ◽  
Vast Number ◽  
Signaling Activation

SUMMARYNeural stem cells (NSCs) and their progeny reside in specialized niches in the adult mammalian brain where they generate new neurons and glia throughout life. Adult NSCs of the ventricular-subventricular zone (V-SVZ) are prone to rapid exhaustion; thus timely, context-dependent neurogenesis demands adaptive signaling among the vast number of neighboring progenitors nestled between the ventricular surface and nearby blood vessels. To dissect adult neuronal lineage progression and regulation, we profiled >56,000 V-SVZ and olfactory bulb (OB) cells by single-cell RNA-sequencing (scRNA-seq). Our analyses revealed the diversity of V-SVZ-derived OB neurons, the temporal dynamics of lineage progression, and a key intermediate NSC population enriched for expression of Notum, which encodes a secreted WNT antagonist. Single Cell Optical Phenotyping and Expression (SCOPE-seq), a technology linking live cell imaging with scRNA-seq, uncovered dynamic control of cell size concomitant with NSC differentiation with Notum+ NSCs at a critical size poised for cell division, and a preference of NOTUM surface binding to neuronal precursors with active WNT signaling. Finally, in vivo pharmacological inhibition of NOTUM significantly expanded neuronal precursor pools in the V-SVZ. Our findings highlight a critical regulatory state during NSC activation marked by NOTUM, a secreted enzyme that ensures efficient neurogenesis by preventing WNT signaling activation in NSC progeny.

scholarly journals Single-cell profiling reveals an endothelium-mediated immunomodulatory pathway in the eye choroid

2020 ◽  
Vol 217 (6) ◽  
Author(s):  
Guillermo L. Lehmann ◽  
Christin Hanke-Gogokhia ◽  
Yang Hu ◽  
Rohan Bareja ◽  
Zelda Salfati ◽  
...  
Keyword(s):  
Single Cell ◽  
Hedgehog Signaling ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Vascular Diseases ◽  
Cell Types ◽  
Significant Loss ◽  
Support Functions ◽  
Downstream Target

The activity and survival of retinal photoreceptors depend on support functions performed by the retinal pigment epithelium (RPE) and on oxygen and nutrients delivered by blood vessels in the underlying choroid. By combining single-cell and bulk RNA sequencing, we categorized mouse RPE/choroid cell types and characterized the tissue-specific transcriptomic features of choroidal endothelial cells. We found that choroidal endothelium adjacent to the RPE expresses high levels of Indian Hedgehog and identified its downstream target as stromal GLI1+ mesenchymal stem cell–like cells. In vivo genetic impairment of Hedgehog signaling induced significant loss of choroidal mast cells, as well as an altered inflammatory response and exacerbated visual function defects after retinal damage. Our studies reveal the cellular and molecular landscape of adult RPE/choroid and uncover a Hedgehog-regulated choroidal immunomodulatory signaling circuit. These results open new avenues for the study and treatment of retinal vascular diseases and choroid-related inflammatory blinding disorders.

scholarly journals Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Kristen L Wells ◽  
Corey N Miller ◽  
Andreas R Gschwind ◽  
Wu Wei ◽  
Jonah D Phipps ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Critical Role ◽  
Lineage Tracing ◽  
Thymic Epithelial Cells ◽  
In Vivo Models ◽  
Single Cell Rna Sequencing ◽  
Medullary Thymic Epithelial Cells

Medullary thymic epithelial cells (mTECs) play a critical role in central immune tolerance by mediating negative selection of autoreactive T cells through the collective expression of the peripheral self-antigen compartment, including tissue-specific antigens (TSAs). Recent work has shown that gene-expression patterns within the mTEC compartment are heterogenous and include multiple differentiated cell states. To further define mTEC development and medullary epithelial lineage relationships, we combined lineage tracing and recovery from transient in vivo mTEC ablation with single-cell RNA-sequencing in Mus musculus. The combination of bioinformatic and experimental approaches revealed a non-stem transit-amplifying population of cycling mTECs that preceded Aire expression. We propose a branching model of mTEC development wherein a heterogeneous pool of transit-amplifying cells gives rise to Aire- and Ccl21a-expressing mTEC subsets. We further use experimental techniques to show that within the Aire-expressing developmental branch, TSA expression peaked as Aire expression decreased, implying Aire expression must be established before TSA expression can occur. Collectively, these data provide a roadmap of mTEC development and demonstrate the power of combinatorial approaches leveraging both in vivo models and high-dimensional datasets.

scholarly journals The embryonic node functions as an instructive stem cell niche

2020 ◽  
Author(s):  
Tatiana Solovieva ◽  
Hui-Chun Lu ◽  
Adam Moverley ◽  
Nicolas Plachta ◽  
Claudio D. Stern
Keyword(s):  
Stem Cell ◽  
Single Cell ◽  
Stem Cell Niche ◽  
Posterior Part ◽  
Growth Zone ◽  
The Body ◽  
Phase Cell ◽  
Cell Behaviour ◽  
Cell Niche

In warm-blooded vertebrate embryos (mammals and birds), the body forms from a growth zone at the tail end. Hensen’s node, a region which induces and patterns the neural axis is located within this growth zone. The node also contains the precursors of neural, mesodermal and endodermal structures along the midline and has been suggested to contain a small population of resident stem cells. However, it is unknown whether the rest of the node constitutes an instructive stem cell niche, specifying stem cell behaviour. Here we combine transplantation of a single cell in vivo with single-cell mRNA sequencing in the chick and show that when made to enter the node, non-node-progenitor cells become resident and gain stem cell behaviour. These cells preferentially express G2/M phase cell-cycle related genes and are concentrated in posterior sub-regions of the node. The posterior part of the node therefore behaves as an instructive stem cell niche. These results demonstrate a new function for the vertebrate node during development.

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