Creation of X-linked Alport syndrome rat model with Col4a5 deficiency

Masumi Namba,Tomoe Kobayashi,Takayuki Koyano,Takuo Hirose,Masaki Fukushima,Mayumi Kohno,Makoto Matsuyama

doi:10.1038/s41598-021-00354-y

Abstract

Alport syndrome is an inherited chronic human kidney disease, characterized by glomerular basement membrane abnormalities. This disease is caused by mutations in COL4A3, COL4A4, or COL4A5 gene. The knockout mice for Col4α3, Col4α4, and Col4α5 are developed and well characterized for the study of Alport syndrome. However, disease progression and effects of pharmacological therapy depend on the genetic variability. This model was reliable only to mouse. In this study, we created a novel Alport syndrome rat model utilizing the rGONAD technology, which generated rat with a deletion of the Col4α5 gene. Col4α5 deficient rats showed hematuria, proteinuria, high levels of BUN, Cre, and then died at 18 to 28 weeks of age (Hemizygous mutant males). Histological and ultrastructural analyses displayed the abnormalities including parietal cell hyperplasia, mesangial sclerosis, and interstitial fibrosis. Then, we demonstrated that α3/α4/α5 (IV) and α5/α5/α6 (IV) chains of type IV collagen disrupted in Col4α5 deficient rats. Thus, Col4α5 mutant rat is a reliable candidate for the Alport syndrome model for underlying the mechanism of kidney diseases and further identifying potential therapeutic targets for human renal diseases.

Highlights

Alport syndrome is an inherited chronic human kidney disease, characterized by glomerular basement membrane abnormalities
Alport syndrome is caused by mutations in COL4A3, COL4A4, or COL4A5 gene encoding type IV collagen α3, α4, or α5 chains[5]
The mutation was verified by PCR followed by DNA sequencing, resulting to be expressed only 9 amino acids of COL4A5 at its N-terminus (Supplementary Fig. 1b,c)

Summary

Introduction

Alport syndrome is an inherited chronic human kidney disease, characterized by glomerular basement membrane abnormalities. This disease is caused by mutations in COL4A3, COL4A4, or COL4A5 gene. Disease progression and effects of pharmacological therapy depend on the genetic variability. This model was reliable only to mouse. Col4α5 mutant rat is a reliable candidate for the Alport syndrome model for underlying the mechanism of kidney diseases and further identifying potential therapeutic targets for human renal diseases. Alport syndrome is caused by mutations in COL4A3, COL4A4, or COL4A5 gene encoding type IV collagen α3, α4, or α5 chains[5]. The recent CRISPR/Cas[9] system is the simplest

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Results

Conclusion