Targeted next-generation sequencing for detection of PIK3CA mutations in archival tissues from patients with Klippel-Trenaunay syndrome in an Asian population : List the full names and institutional addresses for all authors.

Capillary malformations High-throughput nucleotide sequencing Klippel–Trenaunay Syndrome Limb hypertrophy Lymphatic abnormalities PIK3CA-related overgrowth spectrum Phosphatidylinositol 3-Kinase Segmental hypertrophy Vascular malformations Venous malformations

Journal

Orphanet journal of rare diseases
ISSN: 1750-1172
Titre abrégé: Orphanet J Rare Dis
Pays: England
ID NLM: 101266602

Informations de publication

Date de publication:
04 09 2023
Historique:
received: 19 04 2023
accepted: 26 08 2023
medline: 6 9 2023
pubmed: 5 9 2023
entrez: 4 9 2023
Statut: epublish

Résumé

Klippel-Trenaunay syndrome (KTS) is a rare slow-flow combined vascular malformation with limb hypertrophy. KTS is thought to lie on the PIK3CA-related overgrowth spectrum, but reports are limited. PIK3CA encodes p110α, a catalytic subunit of phosphatidylinositol 3-kinase (PI3K) that plays an essential role in the PI3K/AKT/mammalian target of rapamycin (mTOR) signaling pathway. We aimed to demonstrate the clinical utility of targeted next-generation sequencing (NGS) in identifying PIK3CA mosaicism in archival formalin-fixed paraffin-embedded (FFPE) tissues from patients with KTS. Participants were 9 female and 5 male patients with KTS diagnosed as capillaro-venous malformation (CVM) or capillaro-lymphatico-venous malformation (CLVM). Median age at resection was 14 years (range, 5-57 years). Median archival period before DNA extraction from FFPE tissues was 5.4 years (range, 3-7 years). NGS-based sequencing of PIK3CA achieved an amplicon mean coverage of 119,000x. PIK3CA missense mutations were found in 12 of 14 patients (85.7%; 6/8 CVM and 6/6 CLVM), with 8 patients showing the hotspot variants E542K, E545K, H1047R, and H1047L. The non-hotspot PIK3CA variants C420R, Q546K, and Q546R were identified in 4 patients. Overall, the mean variant allele frequency for identified PIK3CA variants was 6.9% (range, 1.6-17.4%). All patients with geographic capillary malformation, histopathological lymphatic malformation or macrodactyly of the foot had PIK3CA variants. No genotype-phenotype association between hotspot and non-hotspot PIK3CA variants was found. Histologically, the vessels and adipose tissues of the lesions showed phosphorylation of the proteins in the PI3K/AKT/mTOR signaling pathway, including p-AKT, p-mTOR, and p-4EBP1. The PI3K/AKT/mTOR pathway in mesenchymal tissues was activated in patients with KTS. Amplicon-based targeted NGS could identify low-level mosaicism from low-input DNA extracted from FFPE tissues, potentially providing a diagnostic option for personalized medicine with inhibitors of the PI3K/AKT/mTOR signaling pathway.

Sections du résumé

BACKGROUND
Klippel-Trenaunay syndrome (KTS) is a rare slow-flow combined vascular malformation with limb hypertrophy. KTS is thought to lie on the PIK3CA-related overgrowth spectrum, but reports are limited. PIK3CA encodes p110α, a catalytic subunit of phosphatidylinositol 3-kinase (PI3K) that plays an essential role in the PI3K/AKT/mammalian target of rapamycin (mTOR) signaling pathway. We aimed to demonstrate the clinical utility of targeted next-generation sequencing (NGS) in identifying PIK3CA mosaicism in archival formalin-fixed paraffin-embedded (FFPE) tissues from patients with KTS.
RESULTS
Participants were 9 female and 5 male patients with KTS diagnosed as capillaro-venous malformation (CVM) or capillaro-lymphatico-venous malformation (CLVM). Median age at resection was 14 years (range, 5-57 years). Median archival period before DNA extraction from FFPE tissues was 5.4 years (range, 3-7 years). NGS-based sequencing of PIK3CA achieved an amplicon mean coverage of 119,000x. PIK3CA missense mutations were found in 12 of 14 patients (85.7%; 6/8 CVM and 6/6 CLVM), with 8 patients showing the hotspot variants E542K, E545K, H1047R, and H1047L. The non-hotspot PIK3CA variants C420R, Q546K, and Q546R were identified in 4 patients. Overall, the mean variant allele frequency for identified PIK3CA variants was 6.9% (range, 1.6-17.4%). All patients with geographic capillary malformation, histopathological lymphatic malformation or macrodactyly of the foot had PIK3CA variants. No genotype-phenotype association between hotspot and non-hotspot PIK3CA variants was found. Histologically, the vessels and adipose tissues of the lesions showed phosphorylation of the proteins in the PI3K/AKT/mTOR signaling pathway, including p-AKT, p-mTOR, and p-4EBP1.
CONCLUSIONS
The PI3K/AKT/mTOR pathway in mesenchymal tissues was activated in patients with KTS. Amplicon-based targeted NGS could identify low-level mosaicism from low-input DNA extracted from FFPE tissues, potentially providing a diagnostic option for personalized medicine with inhibitors of the PI3K/AKT/mTOR signaling pathway.

Identifiants

pubmed: 37667289
doi: 10.1186/s13023-023-02893-1
pii: 10.1186/s13023-023-02893-1
pmc: PMC10478188
doi:

Substances chimiques

Class I Phosphatidylinositol 3-Kinases EC 2.7.1.137
Phosphatidylinositol 3-Kinases EC 2.7.1.-
PIK3CA protein, human EC 2.7.1.137
Proto-Oncogene Proteins c-akt EC 2.7.11.1
TOR Serine-Threonine Kinases EC 2.7.11.1

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

270

Informations de copyright

© 2023. Institut National de la Santé et de la Recherche Médicale (INSERM).

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Auteurs

Yuki Sasaki (Y)

Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, 060-8638, Japan.
Center for Vascular Anomalies, Department of Plastic and Reconstructive Surgery, Tonan Hospital, Hokkaido, Japan.

Kosuke Ishikawa (K)

Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, 060-8638, Japan. kishikawa-hok@umin.ac.jp.
Center for Vascular Anomalies, Department of Plastic and Reconstructive Surgery, Tonan Hospital, Hokkaido, Japan. kishikawa-hok@umin.ac.jp.

Kanako C Hatanaka (KC)

Center for Development of Advanced Diagnostics, Institute of Health Science Innovation for Medical Care, Hokkaido University Hospital, Hokkaido, Japan.

Yumiko Oyamada (Y)

Department of Diagnostic Pathology, Tonan Hospital, Hokkaido, Japan.

Yusuke Sakuhara (Y)

Department of Diagnostic and Interventional Radiology, Tonan Hospital, Hokkaido, Japan.

Tadashi Shimizu (T)

Department of Diagnostic and Interventional Radiology, Tonan Hospital, Hokkaido, Japan.

Tatsuro Saito (T)

Research Division of Genome Companion Diagnostics, Hokkaido University Hospital, Hokkaido, Japan.
Riken Genesis Co., Ltd, Tokyo, Japan.

Naoki Murao (N)

Center for Vascular Anomalies, Department of Plastic and Reconstructive Surgery, Tonan Hospital, Hokkaido, Japan.

Tomohiro Onodera (T)

Department of Orthopedic Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Hokkaido, Japan.

Takahiro Miura (T)

Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, 060-8638, Japan.

Taku Maeda (T)

Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, 060-8638, Japan.

Emi Funayama (E)

Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, 060-8638, Japan.

Yutaka Hatanaka (Y)

Center for Development of Advanced Diagnostics, Institute of Health Science Innovation for Medical Care, Hokkaido University Hospital, Hokkaido, Japan.
Research Division of Genome Companion Diagnostics, Hokkaido University Hospital, Hokkaido, Japan.

Yuhei Yamamoto (Y)

Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo, 060-8638, Japan.

Satoru Sasaki (S)

Center for Vascular Anomalies, Department of Plastic and Reconstructive Surgery, Tonan Hospital, Hokkaido, Japan.

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