Preliminary study on pathogenic mechanism of first Chinese family with PNKD

Feng Chen; Shaohui Zhang; Tinghong Liu; Liu Yuan; Yangshuo Wang; Guojun Zhang; Shuli Liang

doi:10.1515/tnsci-2022-0222

Preliminary study on pathogenic mechanism of first Chinese family with PNKD

Transl Neurosci. 2022 Jun 9;13(1):125-133. doi: 10.1515/tnsci-2022-0222. eCollection 2022 Jan 1.

Authors

Feng Chen¹, Shaohui Zhang², Tinghong Liu^{3

4}, Liu Yuan¹, Yangshuo Wang¹, Guojun Zhang^{3

4}, Shuli Liang^{3

4}

Affiliations

¹ Functional Neurosurgery Department, National Children's Health Center of China, Beijing Children's Hospital, Capital Medical University, No. 56, Nanlishi Road, Xicheng District, Beijing, 100045, China.
² Neurosurgery Department, PLA General Hospital, No. 28 Fuxing Road, Haidian District, Beijing, 100853, China.
³ Functional Neurosurgery Department, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing, China, No. 56, Nanlishi Road, Xicheng District, Beijing, 100045, China.
⁴ Key Laboratory of Major Diseases in Children, Ministry of Education, Beijing, China, No. 56, Nanlishi Road, Xicheng District, Beijing, 100045, China.

Abstract

Background: The first Chinese family with paroxysmal non-kinesigenic dystonia (PNKD) was confirmed to harbour a PNKD mutation. However, the pathogenic mechanism of the PNKD-causing gene mutation was unclear.

Methods: Wild-type and mutant PNKD-L plasmids were prepared and transfected into the C6 cell line to study the distribution and stability of PNKD protein in C6 cells and its effect on the glutathione content. The blood and cerebrospinal fluid (CSF) of 3 PNKD patients and 3 healthy controls were collected. The differentially expressed proteins were identified using isobaric tags for relative and absolute quantitation. Furthermore, Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses were performed, and the protein-protein interaction network was constructed.

Results: Wild-type PNKD protein was mainly distributed in the membranes, whereas mutant PNKD protein was distributed throughout the C6 cells. After transfection with mutant PNKD-L plasmid, the glutathione content decreased significantly in C6 cells; the stability of the mutant PNKD protein was significantly low. There were 172 and 163 differentially expressed proteins in CSF and plasma, respectively, of PNKD patients and healthy controls. For these proteins, blood microparticle and complex activation (classical pathway) were the common GO enrichment term, and complex and coordination cascade pathway were the common KEGG enrichment pathway. Recombinant mothers against decapentaplegic homolog 4 (SMAD4) was one of the differentially expressed proteins; it exhibited a relationship with the aforementioned enrichment GO terms and KEGG pathway.

Conclusion: PNKD protein was mainly distributed in cell membranes. PNKD-L mutation affected subcellular localisation, PNKD protein stability, and glutathione content. SMAD4 was found to be a potential biomarker for PNKD diagnosis.

Keywords: gene function; gene mutation; isobaric tags for relative and absolute quantitation; paroxysmal non-kinesigenic dystonia.