EIF4A3 deficient human iPSCs and mouse models demonstrate neural crest defects that underlie Richieri-Costa-Pereira syndrome

Abstract

Biallelic loss-of-function mutations in the RNA-binding protein EIF4A3 cause Richieri-Costa-Pereira syndrome (RCPS), an autosomal recessive condition mainly characterized by craniofacial and limb malformations. However, the pathogenic cellular mechanisms responsible for this syndrome are entirely unknown. Here, we used two complementary approaches, patient-derived induced pluripotent stem cells (iPSCs) and conditional Eif4a3 mouse models, to demonstrate that defective neural crest cell (NCC) development explains RCPS craniofacial abnormalities. RCPS iNCCs have decreased migratory capacity, a distinct phenotype relative to other craniofacial disorders. Eif4a3 haploinsufficient embryos presented altered mandibular process fusion and micrognathia, thus recapitulating the most penetrant phenotypes of the syndrome. These defects were evident in either ubiquitous or NCC-specific Eif4a3 haploinsufficient animals, demonstrating an autonomous requirement of Eif4a3 in NCCs. Notably, RCPS NCC-derived mesenchymal stem-like cells (nMSCs) showed premature bone differentiation, a phenotype paralleled by premature clavicle ossification in Eif4a3 haploinsufficient embryos. Likewise, nMSCs presented compromised in vitro chondrogenesis, and Meckel's cartilage was underdeveloped in vivo. These findings indicate novel and essential requirements of EIF4A3 for NCC migration and osteochondrogenic differentiation during craniofacial development. Altogether, complementary use of iPSCs and mouse models pinpoint unique cellular mechanisms by which EIF4A3 mutation causes RCPS, and provide a paradigm to study craniofacial disorders.

Effect of knockdown of eIF4A1 by shRNA on the biological characteristics of glioma cells

“…真核生物蛋白翻译过程需要多个因子的参与 [5] . ; eIF4A3与胚胎的早期发育相关 [7] . 研究发 现, eIF4B和eIF4E的表达能够增强eIF4A1的活性 [8] , 而 肿瘤抑制子PDCD4能抑制其功能 [9] .…”

Investigating the pathogenic mechanism underlying Richieri-Costa-Pereira syndrome

“…with the typical immunophenotype, which gave rise to osteoblasts, chondrocytes and adipocytes (Miller et al 2017). Taken together, these data demonstrated that our strategy efficiently yielded disease-relevant cell types, which allowed us to assess cellular and molecular phenotypes associated with RCPS in vitro.…”

“…Despite presenting phenotypes comparable to humans, whose EIF4A3 levels are varied depending on cell type, Eif4a3 haploinsufficient mice also displayed severe phenotypes not so far described in humans (Miller et al 2017). These phenotypic differences could be due to Cre efficiency or because humans are genetically diverse whereas mice are inbred, which exposes phenotypes that are normally masked in humans.…”

“…The cellular mechanisms by which reduced EIF4A3 causes RCPS have been elucidated in chapter III (Miller et al 2017 We provided evidence that impaired NCC migratory capacity is a key cellular dysfunction involved in RCPS pathogenesis. Delayed migration at early developmental stages can reduce mesenchymal progenitors populating the pharyngeal arches and subsequently impair cartilage and bone formation leading to abnormal or absence of mandibular fusion, which is amongst the most relevant RCPS phenotypes.…”

“…In view of the limited access to human embryonic tissues and the fact that animal embryos may not completely recapitulate human embryonic development (Thyagarajan et al 2003;de Bakker et al 2016;, human iPSC-based strategies have been largely used as an alternative approach to study NCC biology in pathological conditions. The available methods for generating NCCs rely on BMP pathway modulation (Jiang et al 2009;Lee et al 2010;Pomp et al 2005;Lee et al 2007;Curchoe et al 2010) and may or may not be combined with WNT pathway activation Miller et al 2017;Menendez et al 2011;Noisa et al 2014;Huang et al 2016;Liu et al 2012;Lee et al 2009;Mica et al 2013;Kreitzer et al 2013;Leung et al 2016;Thier et al 2019;Tchieu et al 2017). However, many of these methods lack a NPB stage or are unable to fully convert neuroepithelial cell populations into NCCs, potentially hindering investigation of the various stages of neural crest specification.…”