Abstract
The pattern of X-chromosome inactivation (XCI) can affect the clinical severity of X-linked disorders in females. XCI pattern analysis has been conducted mainly by HUMARA assay, a polymerase chain reaction-based assay using a methylation-sensitive restriction enzyme. However, this assay examines the XCI ratio of the androgen receptor gene at the genomic DNA level and does not reflect the ratio of either targeted gene directly or at the mRNA level. Here, we report four females with Dent disease, and we clarified the correlation between XCI and female cases of Dent disease using not only HUMARA assay but also a novel analytical method by RNA sequencing. We constructed genetic analysis for 4 female cases showing high level of urinary low-molecular-weight proteinuria and their parents. Their XCI pattern was analyzed by both HUMARA assay and an ultra-deep targeted RNA sequencing of the CLCN5 gene using genomic DNA and mRNA extracted from both leukocytes and urine sediment. All four cases possessed pathogenic variants of the CLCN5 gene. XCI analysis revealed skewed XCI in only two cases, while the other two showed random XCI. All assay results of HUMARA and targeted RNA sequencing in both leukocytes and urinary sediment were clearly identical in all four cases. We developed a novel XCI analytical assay of ultra-deep targeted RNA sequencing and revealed that skewed XCI explains the mechanism of onset of female Dent disease in only half of such cases.
| Original language | English |
|---|---|
| Pages (from-to) | 589-595 |
| Number of pages | 7 |
| Journal | Journal of Human Genetics |
| Volume | 63 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 01-05-2018 |
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All Science Journal Classification (ASJC) codes
- Genetics
- Genetics(clinical)
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Development of ultra-deep targeted RNA sequencing for analyzing X-chromosome inactivation in female Dent disease. / Minamikawa, Shogo; Nozu, Kandai; Nozu, Yoshimi; Yamamura, Tomohiko; Ikeda, Mariko; Nakanishi, Keita; Fujimura, Junya; Horinouchi, Tomoko; Shima, Yuko; Nakanishi, Koichi; Hattori, Masuji; Kanda, Kyoko; Tanaka, Ryojiro; Morisada, Naoya; Nagano, China; Sakakibara, Nana; Nagase, Hiroaki; Morioka, Ichiro; Kaito, Hiroshi; Iijima, Kazumoto.
In: Journal of Human Genetics, Vol. 63, No. 5, 01.05.2018, p. 589-595.Research output: Contribution to journal › Article
TY - JOUR
T1 - Development of ultra-deep targeted RNA sequencing for analyzing X-chromosome inactivation in female Dent disease
AU - Minamikawa, Shogo
AU - Nozu, Kandai
AU - Nozu, Yoshimi
AU - Yamamura, Tomohiko
AU - Ikeda, Mariko
AU - Nakanishi, Keita
AU - Fujimura, Junya
AU - Horinouchi, Tomoko
AU - Shima, Yuko
AU - Nakanishi, Koichi
AU - Hattori, Masuji
AU - Kanda, Kyoko
AU - Tanaka, Ryojiro
AU - Morisada, Naoya
AU - Nagano, China
AU - Sakakibara, Nana
AU - Nagase, Hiroaki
AU - Morioka, Ichiro
AU - Kaito, Hiroshi
AU - Iijima, Kazumoto
PY - 2018/5/1
Y1 - 2018/5/1
N2 - The pattern of X-chromosome inactivation (XCI) can affect the clinical severity of X-linked disorders in females. XCI pattern analysis has been conducted mainly by HUMARA assay, a polymerase chain reaction-based assay using a methylation-sensitive restriction enzyme. However, this assay examines the XCI ratio of the androgen receptor gene at the genomic DNA level and does not reflect the ratio of either targeted gene directly or at the mRNA level. Here, we report four females with Dent disease, and we clarified the correlation between XCI and female cases of Dent disease using not only HUMARA assay but also a novel analytical method by RNA sequencing. We constructed genetic analysis for 4 female cases showing high level of urinary low-molecular-weight proteinuria and their parents. Their XCI pattern was analyzed by both HUMARA assay and an ultra-deep targeted RNA sequencing of the CLCN5 gene using genomic DNA and mRNA extracted from both leukocytes and urine sediment. All four cases possessed pathogenic variants of the CLCN5 gene. XCI analysis revealed skewed XCI in only two cases, while the other two showed random XCI. All assay results of HUMARA and targeted RNA sequencing in both leukocytes and urinary sediment were clearly identical in all four cases. We developed a novel XCI analytical assay of ultra-deep targeted RNA sequencing and revealed that skewed XCI explains the mechanism of onset of female Dent disease in only half of such cases.
AB - The pattern of X-chromosome inactivation (XCI) can affect the clinical severity of X-linked disorders in females. XCI pattern analysis has been conducted mainly by HUMARA assay, a polymerase chain reaction-based assay using a methylation-sensitive restriction enzyme. However, this assay examines the XCI ratio of the androgen receptor gene at the genomic DNA level and does not reflect the ratio of either targeted gene directly or at the mRNA level. Here, we report four females with Dent disease, and we clarified the correlation between XCI and female cases of Dent disease using not only HUMARA assay but also a novel analytical method by RNA sequencing. We constructed genetic analysis for 4 female cases showing high level of urinary low-molecular-weight proteinuria and their parents. Their XCI pattern was analyzed by both HUMARA assay and an ultra-deep targeted RNA sequencing of the CLCN5 gene using genomic DNA and mRNA extracted from both leukocytes and urine sediment. All four cases possessed pathogenic variants of the CLCN5 gene. XCI analysis revealed skewed XCI in only two cases, while the other two showed random XCI. All assay results of HUMARA and targeted RNA sequencing in both leukocytes and urinary sediment were clearly identical in all four cases. We developed a novel XCI analytical assay of ultra-deep targeted RNA sequencing and revealed that skewed XCI explains the mechanism of onset of female Dent disease in only half of such cases.
UR - http://www.scopus.com/inward/record.url?scp=85042190561&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85042190561&partnerID=8YFLogxK
U2 - 10.1038/s10038-018-0415-1
DO - 10.1038/s10038-018-0415-1
M3 - Article
AN - SCOPUS:85042190561
VL - 63
SP - 589
EP - 595
JO - Journal of Human Genetics
JF - Journal of Human Genetics
SN - 1434-5161
IS - 5
ER -