eISSN: 2299-0046
ISSN: 1642-395X
Advances in Dermatology and Allergology/Postępy Dermatologii i Alergologii
Current issue Archive Manuscripts accepted About the journal Editorial board Reviewers Abstracting and indexing Subscription Contact Instructions for authors Ethical standards and procedures
Editorial System
Submit your Manuscript
SCImago Journal & Country Rank
vol. 40
Original paper

The role of gene encoding collagen secretion protein (SERPINH1) in the pathogenesis of a hypermobile type of Ehlers-Danlos syndrome

Anna Junkiert-Czarnecka
Maria Pilarska-Deltow
Aneta Bąk
Marta Heise
Olga Haus

Department of Clinical Genetics, Faculty of Medicine, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Poland
Adv Dermatol Allergol 2023; XL (1): 102-106
Online publish date: 2023/01/13
Article file
- The role of gene.pdf  [0.17 MB]
Get citation


Hypermobile Ehlers-Danlos syndrome (hEDS, OMIM#130020) is a non-inflammatory, autosomal dominant (for most cases) connective tissue disorder. According to the 2017 EDS International Classification, it is perhaps the most common hereditary connective tissue disorder with a prevalence estimated at 1/5000 [1]. Clinical manifestations include joint hypermobility with recurrent joint dislocations, chronic, widespread pain, mild hyperextensibility of the skin, recurrent or multiple abdominal hernias, atrophic scarring, gastrointestinal disorders, cardiovascular, immune, and nervous system dysfunctions, and in women a high-risk of premature delivery [2]. hEDS, unlike other types of EDS, has no known genetic aetiology, so diagnosis is conducted based on a person’s medical history, a physical examination, and exclusion of other types of EDS after genetic tests. Due to many clinical features that are common with other types of EDS (especially with the EDS classical type) and other non-inflammatory connective tissue disorders, proper recognition of this condition is difficult in many patients. Some reports point to the TNXB gene as an hEDS genetic factor [3, 4]. However, the fact that mutations in TNXB were detected only in a small percentage of hEDS as well as their recessive inheritance patterns suggests that TNXB is not the main hEDS-related gene. Investigations with the application of a next-generation sequencing (NGS) connective tissue panel, performed in hEDS patients, also do not identify disorder-associated genes. In several studies, new mutations were detected in well-known genes encoding extracellular matrix proteins, such as COL5A1, COL5A2, and COL3A1 [57].

In an investigation by Chiarelli et al., expression profiling of some ECM proteins in hEDSpatients’ fibroblasts was done. The study showed that ECM proteins, especially collagen type I, III, and V, as well as fibrillin and fibronectin, were detectable only in patients’ cell cytoplasm, whereas in intercellular spaces they were either not visible or only a few thin fibrils were present. Meanwhile, in healthy control cells, proteins were visible inside and outside the cells [8]. The cause of the dysfunction of ECM protein transport from cytoplasm to the intracellular space in hEDS patients is unknown. In the pathway of ECM protein transport, many molecules are involved in several stages. One of these proteins is heat-shock protein 47 (HSP47) encoded by the SERPINH1 gene.

Collagen, the most abundant human protein, is a major component of the extracellular matrix (ECM) in tissues such as skin, bone, ligaments, tendons, arteries, and veins, as well as the gastrointestinal tract and respiratory system. Collagen biosynthesis is a complex, multistage process incorporating many molecules. It takes place in the rough endoplasmic reticulum (rER), and then the collagen molecule is transported via the Golgi apparatus to the extracellular space. All human proteins, including collagen molecules, require proper protein folding, maturation, and secretion processes. In the lumen of the rER collagen molecules undergo a series of posttranslational modifications, e.g. hydroxylation of propyl and lysyl residues by propyl 4-hydroxylases (P4H), propyl 3-hydroxylases (P3H), and lysyl hydroxylases (LH), respectively, and glycosylation of hydroxylysyl residues by glycosyltransferases. Other essential proteins involved in collagen folding and maturation are chaperone proteins such as HSP47 [9]. Heat-shock protein (HSP) 47 is a rough endoplasmic reticulum resident molecular chaperone, belonging to the serpin (serine protease inhibitor) superfamily [9, 10]. HSP47 is essential for the proper assembly of procollagen in the rER, which is eventually transported across the Golgi apparatus to the extracellular space (Figure 1). HSP47 was shown to directly bind to types I, II, III, IV, and V of collagen. It has been proposed that HSP47 serves 2 functions as a molecular chaperone in the collagen synthetic pathway: inhibition of local unfolding procollagen, and binding and stabilization of the triple-helical form of procollagen, and prevention of procollagen aggregation in the rER (prevention of rER-stress) [11, 12].

Figure 1

Collagen folding and HSP47. HSP47 binds to triple-helical procollagen in the rER and dissociates in the Golgi apparatus [9]. Figure created by BioRender


HSP47 is encoded by the SERPINH1 gene consisting of 5 exons. It has never been examined in hypermobile-type Ehlers-Danlos syndrome (hEDS) patients. The role of HSP47 has been evaluated in Osteogenesis Imperfecta (OI), but never in other connective tissue disorders. However, many hEDS patients share the phenotype with OI patients in whom the disorder develops based on COL1A1 variants [1317]. Ishida et al. investigated biallelic knockout mice (HSP47-/-) cells for the role of HSP47 in type I collagen maturation and secretion from the rough endoplasmic reticulum to the Golgi apparatus. The secretion rate of this type of collagen was reduced in comparison to mouse cells HSP47+/+. They also noticed that in HSP47-/- cells, collagen molecules accumulated in the rER, compared to HSP47+/+ cells, in which such accumulation was not detected [18].


The aim of the present study was a sequencing analysis of the SERPINH1 gene and an evaluation of the potential role of variants of this gene on the aetiology of the hEDS.

Material and methods

The study group included 100 hEDS patients of Polish origin, women (84) and men (16), aged 17–63 years (median: 31 years). Patients were enrolled in the study by experienced clinical geneticists, according to the 2017 International Classification of the Ehlers-Danlos syndrome diagnostic criteria [1]. Joint hypermobility was evaluated on the Beighton scale. The control group consisted of 100 volunteers from the general Polish population matched by age and sex with the investigated group, who were healthy (including lack of EDS) at the time of the investigation and without a history of EDS in the family.

All hEDS patients or their parents provided informed consent to participate in the study. Consent to publish clinical/genetic data was also obtained from the patients or their parents.

The study was approved by the Ethics Committee of Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Poland (KB485/2013).

The analysis was performed on genomic DNA (gDNA), which was extracted from leukocytes (fibroblasts were not available) by QIAamp DNA Mini Kit (Qiagen, Germany) using standard procedures. In all patients, other types of EDS or other connective tissue disorders were excluded by testing them with NGS technology (Illumina NextSeq 550). The connective tissue disorder customer panel included COL5A1, COL5A2, COL3A1, COL1A1, COL1A2, TNXB, ADAMTS2, PLOD1, FKBP14, ZNF469, PRDM5, B4GALT7, B3GALT6, SLC39A13, CHST14, DSE, COL12A1, C1R, C1S, SEC23A, SEC24D, COL6A1, COL6A2, COL6A3, COL9A1, COL9A2, FBN1, FBN2, FLNA, and FLNB. Copy number variant (CNV) analysis was also performed. Sequencing data were aligned to the hg19 human reference genome. Based on the guidelines of the American College of Medical Genetics and Genomics, a minimum depth coverage of 30X was considered suitable for analysis. All patients negative for the NGS test were analysed for alterations in the SERPINH1 gene. Molecular analysis of the SERPINH1 gene was performed by Sanger sequencing (ABI3130XL) according to standard procedure (primer sequences available upon request). The pathogenicity of detected variants was assessed according to the ACMG guideline by Varsome [19, 20].

Statistical analysis

For statistics, the χ2 test was used.


All patients were tested by NGS multi-gene panel for connective tissue disorders, such as EDS, Osteogenesis Imperfecta, Marfan syndrome, and others. No pathogenic or likely pathogenic variants were detected in any of them. Copy number variants were also not detected.

The next step of the investigation was the sequencing of the SERPINH1 gene by Sanger sequencing.

Among 100 tested patients, 4 different types of missense variants (heterozygote) were detected. All SERPINH1 alterations were classified as benign according to ACMG guidelines (Table 1).

Table 1

Characterization of variants detected in the SERPINH1 gene

NVariantdbSNPFrequency (patients)Frequency (control group)P-valuePathogenicity (according to ACMG)
1c.267G>A, p.Thr89=rs1391274262/1000/1000.49Benign
2c.693C>T, p.Thr231=rs64925736/10030/1000.45Benign
3c.744C>T, p.Asp248=rs617332481/1000/1001.0Benign
4c.1011G>A, p.Leu337=rs58582125/10030/1000.52Benign

No pathogenic or probably pathogenic alterations were found. Variants were located in all exons of the gene, except for exon 1, which is non-coding. Part of exon 2 (from nucleotide number 60) to the beginning of exon 5 (nucleotide number 1159) correspond to the N-terminus part of the protein (Figure 2). All detected variants were found in the N-terminus protein domain, in C-terminus, no variant was located.

Figure 2

Location of the detected variants along the SERPINH1 gene (Figure created by BioRender)


We also noticed that there were differences between the location of variants and patients’ clinical symptoms (severity and intensity of the symptoms, sex, age of onset of the disease), and also family history etc.


Previous molecular analysis of the hypermobile type of Ehlers-Danlos syndrome focused mainly on a wide range of sequencing analyses of genes encoding connective tissue molecules. Despite intensive investigation, genes related to hEDS have not been identified. Some patients with EDS may share symptoms with patients with a mild form of Osteogenesis Imperfecta; furthermore, cases of EDS patients with COL1A1 mutations (a gene strictly connected with OI) are well known [15, 17, 21, 22]. Some phenotype-genotype similarities between EDS and OI were an implication for us to determine the role of another OI gene, SERPINH1, in the molecular background of hEDS [13]. Mutations in the SERPINH1 gene were described in a rare type of OI but have never been analysed in hypermobile Ehlers-Danlos syndrome. In our investigation among 100 hEDS patients, we did not identify pathogenic or probably pathogenic variants. Although only benign variants were detected, which play no role in the pathogenesis of hEDS, we should take into account mechanisms other than gene structure alterations, which may have an impact on collagen and other ECM proteins transport.

We are aware that the gene investigated by us encodes a protein that is only a part of the collagen/ECM secretion pathway, and the potential role of its disrupted distribution may be one of the mechanisms underlying hypermobile Ehlers-Danlos.


The authors would like to thank the patients and their families for participating in the investigation.

This investigation was supported by a grant from National Science Centre Poland (Narodowe Centum Nauki) (2018/29/N/NZ5/00345).

Conflict of interest

The authors declare no conflict of interest.



Malfait F, Francomano C, Byers P, et al. The 2017 international classification of the Ehlers-Danlos syndromes. Am J Med Genet C Semin Med Genet 2017; 175: 8-26.


Tinkle B, Castori M, Berglund B, et al. Hypermobile Ehlers-Danlos syndrome (a.k.a. Ehlers-Danlos syndrome type III and Ehlers-Danlos syndrome hypermobility type): clinical description and natural history. Am J Med Genet C Semin Med Genet 2017; 175: 48-69.


Zweers MC, Bristow J, Steijlen PM, et al. Haploinsufficiency of TNXB is associated with hypermobility type of Ehlers-Danlos syndrome. Am J Hum Genet 2003; 73: 214-7.


Kaufman CS, Butler MG. Mutation in TNXB gene causes moderate to severe Ehlers-Danlos syndrome. World J Med Genet 2016; 6: 17-21.


Weerakkody RA, Vandrovcova J, Kanonidou C, et al. Targeted next-generation sequencing makes new molecular diagnoses and expands genotype–phenotype relationship in Ehlers-Danlos syndrome. Genet Med 2016; 18: 1119-27.


Cortini F, Marinelli B, Romi S, et al. A new COL3A1 mutation in Ehlers-Danlos syndrome vascular type with different phenotypes in the same family. Vasc Endovascular Surg 2017; 51: 141-5.


Zhang W, Han Q, Zhou M, et al. Identification of a missense mutation of COL3A1 in a Chinese family with atypical Ehlers-Danlos syndrome using targeted next-generation sequencing. Mol Med Rep 2017; 15: 936-40.


Chiarelli N, Carini G, Zoppi N, et al. Transcriptome-wide expression profiling in skin fibroblasts of patients with joint hypermobility syndrome/Ehlers-Danlos syndrome hypermobility type. PLoS One 2016; 11: e0161347.


Ito S, Nagata K. Biology of Hsp47 (Serpin H1), a collagen-specific molecular chaperone. Semin Cell Dev Biol 2017; 62: 142-51.


Natsume T, Koide T, Yokota S, et al. Interactions between collagen-binding stress protein HSP47 and collagen. Analysis of kinetic parameters by surface plasmon resonance biosensor. J Biol Chem 1994; 269: 31224-8.


Ishikawa Y, Holden P, Bächinger HP. Heat shock protein 47 and 65-kDa FK506-binding protein weakly but synergistically interact during collagen folding in the endoplasmic reticulum. J Biol Chem 2017; 292: 17216-24.


Widmer C, Gebauer JM, Brunstein E, et al. Molecular basis for the action of the collagen-specific chaperone Hsp47/SERPINH1 and its structure-specific client recognition. Proc Natl Acad Sci USA 2012; 109: 13243-7.


Song Y, Zhao D, Xu X, et al. Novel compound heterozygous mutations in SERPINH1 cause rare autosomal recessive osteogenesis imperfecta type X. Osteoporosis Int 2018; 29: 1389-96.


Ito S, Nagata K. Mutants of collagen-specific molecular chaperone Hsp47 causing osteogenesis imperfecta are structurally unstable with weak binding affinity to collagen. Biochem Biophys Res Commun 2016; 469: 437-42.


Symoens S, Steyaert W, Demuynck L, et al. Tissue-specific mosaicism for a lethal osteogenesis imperfecta COL1A1 mutation causes mild OI/EDS overlap syndrome. Am J Med Genet A 2017; 173: 1047-50.


Mackenroth L, Fischer-Zirnsak B, Egerer J, et al. An overlapping phenotype of osteogenesis imperfecta and Ehlers-Danlos syndrome due to a heterozygous mutation in COL1A1 and biallelic missense variants in TNXB identified by whole exome sequencing. Am J Med Genet A 2016; 170: 1080-5.


Colombi M, Dordoni C, Venturini M, et al. Delineation of Ehlers-Danlos syndrome phenotype due to the c.934C>T, p.(Arg312Cys) mutation in COL1A1: report on a three-generation family without cardiovascular events, and literature review. Am J Med Genet A 2017; 173: 524-30.


Ishida Y, Kubota H, Yamamoto A, et al. Type I collagen in Hsp47-null cells is aggregated in endoplasmic reticulum and deficient in n-propeptide processing and fibrillogenesis. Mol Biol Cell 2006; 17: 2346-55.


Kopanos C, Tsiolkas V, Kouris A, et al. VarSome: the human genomic variant search engine. Bioinformatics 2019; 35: 1978-80.


Richards S, Aziz N, Bale S, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med 2015; 17: 405-24.


Mackenroth L, Fischer-Zirnsak B, Egerer J, et al. An overlapping phenotype of osteogenesis imperfecta and Ehlers-Danlos syndrome due to a heterozygous mutation in COL1A1 and biallelic missense variants in TNXB identified by whole exome sequencing. Am J Med Genet A 2016; 170: 1080-5.


Morabito LA, Allegri AEM, Capra AP, et al. Osteogenesis imperfecta/Ehlers-Danlos overlap syndrome and neuroblastoma – case report and review of literature. Genes 2022; 13: 581.

Copyright: © 2023 Termedia Sp. z o. o. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) License (http://creativecommons.org/licenses/by-nc-sa/4.0/), allowing third parties to copy and redistribute the material in any medium or format and to remix, transform, and build upon the material, provided the original work is properly cited and states its license.
Quick links
© 2023 Termedia Sp. z o.o.
Developed by Bentus.