W. Yan (Nanjing, CN)

Nanjing University

Presenter Of 1 Presentation

Poster Cartilage Imaging and Functional Testing

P019 - Variants in LRP1 Links Triradiate Chondrocytes Differentiation are Responsible for DDH in Humans and Mice

Presentation Topic
Cartilage Imaging and Functional Testing
Date
13.04.2022
Lecture Time
09:30 - 09:30
Room
Exhibition Foyer
Session Name
7.3 - Poster Viewing / Coffee Break / Exhibition
Session Type
Poster Session
Disclosure
No Significant Commercial Relationship

Abstract

Purpose

Developmental dysplasia of the hip (DDH) is one of the most common congenital malformations, and it exhibits autosomal dominant inheritance by unknown mechanisms. The pathological mechanisms of DDH are poorly understood previously, which hampers the development of diagnostic tools and treatments.

Methods and Materials

We conducted whole-exome sequencing and identified 10 distinct variants including missense and splice site mutations of the LRP1 (low density lipoprotein receptor-related protein 1) gene in 2 consanguineous families and 7 unrelated DDH cases from a cohort of 8 consanguineous families and another cohort of 68 unrelated cases DDH-affected Han Chinese child, approximately 13% of the individuals carry novel/rare variants in LRP1. All of the patients harboring LRP1 variants presented a typical DDH phenotype and had complete dislocation of the hips.

We establish a mouse line with Lrp1 gene variants knock-in (KI) via CRISPR-Cas9 System. Micro-CT reconstruction of the acetabular and femoral head of each mice were conducted. Acetabular fossa and femur were rinsed overnight with clean water for hisological staining. The isolation and culture of mice BMSCs from the tibias and femurs of 4-wk-old WT and Lrp1+/- C57/B6 mice were performed. Pellet generate from the BMSCs. Expression level of Lrp1 was specifically knock down in ATDC5 cells using concentrated lentiviruses expressed control-sgRNA or Lrp1-sgRNA.

Results

The timing of triradiate cartilage had been brought forward one or two weeks early in Lrp1R1783W-mutated and Lrp1+/- mice, which stop the acetabulum from growing deeper and wider. Lack of Lrp1 caused a significant decreased chondrogenic ability in vitro. Inhibited autophagy levels and significant up-regulated β-catenin during the chondrogenic induction of mice BMSCs and ADTC5 cell line were detected. Furthermore, we succeeded in rescuing the expression of Col2a1 and Sox9 by treat with 10 or 20 mM PNU in vitro.

Conclusion

Our genetic and protein interaction evidence could open avenues for future studies of DDH.

Collapse

Presenter Of 1 Presentation

Cartilage Imaging and Functional Testing

P019 - Variants in LRP1 Links Triradiate Chondrocytes Differentiation are Responsible for DDH in Humans and Mice

Abstract

Purpose

Developmental dysplasia of the hip (DDH) is one of the most common congenital malformations, and it exhibits autosomal dominant inheritance by unknown mechanisms. The pathological mechanisms of DDH are poorly understood previously, which hampers the development of diagnostic tools and treatments.

Methods and Materials

We conducted whole-exome sequencing and identified 10 distinct variants including missense and splice site mutations of the LRP1 (low density lipoprotein receptor-related protein 1) gene in 2 consanguineous families and 7 unrelated DDH cases from a cohort of 8 consanguineous families and another cohort of 68 unrelated cases DDH-affected Han Chinese child, approximately 13% of the individuals carry novel/rare variants in LRP1. All of the patients harboring LRP1 variants presented a typical DDH phenotype and had complete dislocation of the hips.

We establish a mouse line with Lrp1 gene variants knock-in (KI) via CRISPR-Cas9 System. Micro-CT reconstruction of the acetabular and femoral head of each mice were conducted. Acetabular fossa and femur were rinsed overnight with clean water for hisological staining. The isolation and culture of mice BMSCs from the tibias and femurs of 4-wk-old WT and Lrp1+/- C57/B6 mice were performed. Pellet generate from the BMSCs. Expression level of Lrp1 was specifically knock down in ATDC5 cells using concentrated lentiviruses expressed control-sgRNA or Lrp1-sgRNA.

Results

The timing of triradiate cartilage had been brought forward one or two weeks early in Lrp1R1783W-mutated and Lrp1+/- mice, which stop the acetabulum from growing deeper and wider. Lack of Lrp1 caused a significant decreased chondrogenic ability in vitro. Inhibited autophagy levels and significant up-regulated β-catenin during the chondrogenic induction of mice BMSCs and ADTC5 cell line were detected. Furthermore, we succeeded in rescuing the expression of Col2a1 and Sox9 by treat with 10 or 20 mM PNU in vitro.

Conclusion

Our genetic and protein interaction evidence could open avenues for future studies of DDH.

Collapse