Lineage-specific differences and regulatory networks governing human chondrocyte development
Abstract
To address large gaps in our understanding of the molecular regulation of articular and growth plate cartilage development in humans, we used our directed differentiation approach to generate these distinct cartilage tissues from human embryonic stem cells. The resulting transcriptomic profiles of hESC-derived articular and growth plate chondrocytes were similar to fetal epiphyseal and growth plate chondrocytes, with respect to genes both known and previously unknown to cartilage biology. With the goal to characterize the regulatory landscapes accompanying these respective transcriptomes, we mapped chromatin accessibility in hESC-derived chondrocyte lineages, and mouse embryonic chondrocytes, using ATAC-sequencing. Integration of the expression dataset with the differentially accessible genomic regions revealed lineage-specific gene regulatory networks. We validated functional interactions of two transcription factors (RUNX2 in growth plate chondrocytes and RELA in articular chondrocytes) with their predicted genomic targets. The maps we provide thus represent a framework for probing regulatory interactions governing chondrocyte differentiation. This work constitutes a substantial step towards comprehensive and comparative molecular characterizations of distinct chondrogenic lineages, and sheds new light on human cartilage development and biology.
Data availability
The accession number for the raw ATAC-seq and RNA-seq datasets reported in this paper will be publicly available upon acceptance on GEO under accession GSE195688.
-
Lineage-Specific Differences and Inference of Regulatory Networks Governing Human Chondrocyte DevelopmentNCBI Gene Expression Omnibus, GSE195688.
-
Regulatory constraint and selection during human knee evolution drive modern osteoarthritis riskNCBI Gene Expression Omnibus, GSE122877.
Article and author information
Author details
Funding
NIAMS (R01-AR073821)
- April M Craft
NIAMS (R01-AR070139)
- Terence D Capellini
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Di Chen, Chinese Academy of Sciences, China
Ethics
Animal experimentation: All animal work was performed according to approved institutional animal care and use committee protocols at Harvard University (IACUC 13-04-161).
Human subjects: Human fetal donor samples were collected from the first trimester termination via the University of Washington (UW) Birth Defects Research Laboratory (BRDL) in full compliance with the ethical guidelines of the NIH and with the approval of UW Review Boards for the collection and distribution of human tissue for research, and Harvard University and Boston Children's Hospital for the receipt and use of such materials, and Harvard University and Boston Children's Hospital for the receipt and use of such materials (Capellini: IRB16-1504; Craft: IRB-P00017303). This is not deemed human subjects research.All reported research involving human embryonic stem cells was approved by IRB (IRB-P00017303) and ESCRO (ESCRO-2015.4.24) regulatory bodies at Boston Children's Hospital.
Version history
- Received: May 3, 2022
- Preprint posted: May 25, 2022 (view preprint)
- Accepted: March 14, 2023
- Accepted Manuscript published: March 15, 2023 (version 1)
- Version of Record published: April 3, 2023 (version 2)
Copyright
© 2023, Richard et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 1,373
- views
-
- 253
- downloads
-
- 8
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Further reading
-
- Developmental Biology
- Medicine
From a forward mutagenetic screen to discover mutations associated with obesity, we identified mutations in the Spag7 gene linked to metabolic dysfunction in mice. Here, we show that SPAG7 KO mice are born smaller and develop obesity and glucose intolerance in adulthood. This obesity does not stem from hyperphagia, but a decrease in energy expenditure. The KO animals also display reduced exercise tolerance and muscle function due to impaired mitochondrial function. Furthermore, SPAG7-deficiency in developing embryos leads to intrauterine growth restriction, brought on by placental insufficiency, likely due to abnormal development of the placental junctional zone. This insufficiency leads to loss of SPAG7-deficient fetuses in utero and reduced birth weights of those that survive. We hypothesize that a ‘thrifty phenotype’ is ingrained in SPAG7 KO animals during development that leads to adult obesity. Collectively, these results indicate that SPAG7 is essential for embryonic development and energy homeostasis later in life.
-
- Developmental Biology
- Stem Cells and Regenerative Medicine
Amniogenesis, a process critical for continuation of healthy pregnancy, is triggered in a collection of pluripotent epiblast cells as the human embryo implants. Previous studies have established that bone morphogenetic protein (BMP) signaling is a major driver of this lineage specifying process, but the downstream BMP-dependent transcriptional networks that lead to successful amniogenesis remain to be identified. This is, in part, due to the current lack of a robust and reproducible model system that enables mechanistic investigations exclusively into amniogenesis. Here, we developed an improved model of early amnion specification, using a human pluripotent stem cell-based platform in which the activation of BMP signaling is controlled and synchronous. Uniform amniogenesis is seen within 48 hr after BMP activation, and the resulting cells share transcriptomic characteristics with amnion cells of a gastrulating human embryo. Using detailed time-course transcriptomic analyses, we established a previously uncharacterized BMP-dependent amniotic transcriptional cascade, and identified markers that represent five distinct stages of amnion fate specification; the expression of selected markers was validated in early post-implantation macaque embryos. Moreover, a cohort of factors that could potentially control specific stages of amniogenesis was identified, including the transcription factor TFAP2A. Functionally, we determined that, once amniogenesis is triggered by the BMP pathway, TFAP2A controls the progression of amniogenesis. This work presents a temporally resolved transcriptomic resource for several previously uncharacterized amniogenesis states and demonstrates a critical intermediate role for TFAP2A during amnion fate specification.