Transcriptome analysis of skin fibroblasts with dominant negative COL3A1 mutations provides molecular insights into the etiopathology of vascular Ehlers-Danlos syndrome

May combine two claims.

CDKN2B showed enhanced expression in vEDS cells, supporting microarray data.

“Conversely, CDKN2B, which encodes a cyclin-dependent kinase (CDK) inhibitor, showed an enhanced expression in vEDS cells supporting microarray data (Fig 2A).”

May combine two claims.

Gene expression profiling identified a total of 969 DEGs in vEDS patients' cells compared to controls: 281 up-regulated and 688 down-regulated.

“Gene expression profiling identified a total of 969 DEGs in vEDS patients’ cells when compared to controls: of these, 281 genes were up-regulated, and 688 were down-regulated (S3 Table).”

• Also stated in: § Gene expression profiling · plain

  “The volcano plot depicts all statistically significant 969 DEGs (688 down-regulated, 281 up-regulated) identified in vEDS cells.”

May combine two claims.

Control cells organized HS-GAGs in a rich fibrillar network, whereas in vEDS cells only few fibrils, localized in the extracellular spaces and discrete spots associated with the cell surface, were identified.

“Moreover, control cells organized HS-GAGs in a rich fibrillar network, whereas in vEDS cells only few fibrils, localized in the extracellular spaces and discrete spots associated”

May combine two claims.

IF analyses of COLLIII in present skin fibroblasts with different structural mutations confirmed ECM disorganization and intracellular retention of the aberrant protein.

“IF analyses of COLLIII in the present skin fibroblasts with different structural mutations confirmed the ECM disorganization and intracellular retention of the aberrant protein (Fig 4A).”

May combine two claims.

FBNs were organized in fibrillar networks covering control fibroblasts, whereas FBNs were not assembled into the ECM of vEDS cells, which showed rare and sparse cytoplasmic spots.

“As shown in Fig 4A, FBNs were organized in fibrillar networks covering control fibroblasts, whereas these proteins were not assembled into the ECM of vEDS cells that showed rare and sparse cytoplasmic spots.”

May combine two claims.

FBN2 transcript is strongly down-regulated in vEDS fibroblasts, shown by microarray analysis and confirmed by qPCR.

“This finding is in line with the strong down-regulation of the FBN2 transcript shown by microarray analysis and confirmed by qPCR.”

May combine two claims.

FKBP22 cytoplasmic levels are lower in vEDS patients compared to control cells; WB confirmed this down-regulation demonstrating a 4.1 to 6.4-fold decrease.

“IF of FKBP22 suggested lower cytoplasmic levels of this COLLs-modifying enzyme in patients compared to control cells; WB confirmed this down-regulation demonstrating a 4.1 to 6.4-fold decrease.”

May combine two claims.

Control and vEDS fibroblasts showed comparable levels of PDI, although in mutant cells the IF signals suggested an enlargement of ER vesicles.

“Control and vEDS fibroblasts showed comparable levels of PDI, although in mutant cells the IF signals suggested an enlargement of ER vesicles (Fig 4C).”

May combine two claims.

The most down-regulated genes in vEDS patients' cells were implicated in processes related to cell division, DNA replication, telomere organization, and associated with nucleosome and chromatin assembly.

“The most down-regulated genes in vEDS patients ‘cells were implicated in processes related to cell division, DNA replication, telomere organization, and associated with nucleosome and chromatin assembly.”

May combine two claims.

These mutations act through a dominant negative effect and are associated with the severe clinical spectrum of vEDS, characterized by fragility of soft connective tissues with arterial and organ ruptures.

“through a dominant negative effect are associated with the severe clinical spectrum potentially lethal of vEDS, characterized by fragility of soft connective tissues with arterial and organ ruptures.”

• Also stated in: § Introduction · plain

  “This type of mutations produces a dominant negative effect on the protein, thereby inhibiting extracellular accumulation of mature COLLIII.”

• cites [8] LT Smith, 1997, Mutations in the COL3A1 gene result in the Ehlers-Danlos syndrome type IV and alterations in the size and distribution of the major collagen fibrils of the dermis, J Invest Dermatol sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Predominant expression of COLLIII in walls of blood vessels and hollow organs explains why its reduction due to COL3A1 mutations leads to increased bruising, arterial and bowel rupture, and uterine, cervical, and vaginal fragility during pregnancy and delivery.

“The predominant expression of COLLIII in the walls of blood vessels and in hollow organs explains why its reduction due to COL3A1 mutations leads to increased bruising, arterial and bowel rupture, and uterine, cervical, and vaginal fragility during pregnancy and delivery [4].”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Vascular fragility and abnormal vessel structure cause easy bruising, severe varicosities, arterial dissection or aneurysm formation and eventual arterial or intestinal rupture that can lead to fatal consequences and reduced life span.

“Vascular fragility and abnormal vessel structure cause easy bruising, severe varicosities, arterial dissection or aneurysm formation and eventual arterial or intestinal rupture that can lead to fatal consequences and a reduced life span of patients [4,5].”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

• cites [5] A Busch, 2016, Vascular type Ehlers-Danlos syndrome is associated with platelet dysfunction and low vitamin D serum concentration, Orphanet J Rare Dis sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Quality control was performed by investigating principal component analysis to detect grouping patterns in the samples and to identify the outliers and evaluate whether batch effect significantly affected the data.

“Quality control was performed by investigating principal component analysis to detect grouping patterns in the samples and to identify the outliers and evaluate whether batch effect significantly affected the data.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Heterozygosity for COL3A1 null alleles typically produces a milder clinical phenotype with a significant delayed onset of complications.

“Heterozygosity for COL3A1 null alleles typically produces a milder clinical phenotype with a significant delayed onset of complications [4, 9–12].”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Two available mouse models for vEDS are homozygous or heterozygous nulls, which represent only about 5% or less of all known cases of vEDS patients.

“Currently, there are two available mouse models for vEDS, both are homozygous or heterozygous nulls, which represent only about 5% or less of all known cases of vEDS patients [13–15].”

• cites [13] X Liu, 1997, Type III collagen is crucial for collagen I fibrillogenesis and for normal cardiovascular development, Proc Natl Acad Sci USA sourceNot checked yet

  Not checked in the free run.

• cites [14] TK Cooper, 2010, The haploinsufficient Col3a1 mouse as a model for vascular Ehlers–Danlos syndrome, Vet Pathol sourceNot checked yet

  Not checked in the free run.

• cites [15] LB Smith, 2011, Haploinsufficiency of the murine Col3a1 locus causes aortic dissection: a novel model of the vascular type of Ehlers-Danlos syndrome, Cardiovasc Res sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Homozygotes exhibit a severe perinatal mortality owing to arterial ruptures.

“homozygotes exhibit a severe perinatal mortality owing to arterial ruptures [13].”

• cites [13] X Liu, 1997, Type III collagen is crucial for collagen I fibrillogenesis and for normal cardiovascular development, Proc Natl Acad Sci USA sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Null mutations cause half the normal amount of COLLIII to be secreted.

“These null mutations cause half the normal amount of COLLIII to be secreted [11,12].”

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Mutations leading to premature termination of translation and nonsense-mediated mRNA decay are rare (less than 5%).

“Mutations leading to premature termination of translation and nonsense-mediated mRNA decay are rare (less than 5%).”

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Col3a1+/− mice present a very mild phenotype, as suggested by a low incidence of vascular events.

“While Col3a1+/− mice present a very mild phenotype, as suggested by a low incidence of vascular events”

• cites [13] X Liu, 1997, Type III collagen is crucial for collagen I fibrillogenesis and for normal cardiovascular development, Proc Natl Acad Sci USA sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Substitutions of bulky residues (Arg, Asp, Glu, Val) for glycine residues in the triple helical domain are usually less severe than splice site mutations, but more consequential than substitutions by smaller residues (Ala, Ser, Cys).

“Substitutions of bulky residues (Arg, Asp, Glu, Val) for glycine residues in the triple helical domain are usually less severe than splice site mutations, but more consequential than substitutions by smaller residues (Ala, Ser, Cys).”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Multiple testing correction was applied to control the false-discovery rate (FDR) using the Benjamini-Hochberg (BH) procedure.

“Multiple testing correction was applied to control the false-discovery rate (FDR) using the Benjamini–Hochberg (BH) procedure [20].”

• cites [20] Y Benjamini, 1995, Controlling the false discovery rate: a practical and powerful approach to multiple testing, J R Stat Soc B Stat Methodol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Splice site mutations that lead to exon skipping have the lowest median survival.

“Splice site mutations that lead to exon skipping have the lowest median survival.”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

The nature of the specific COL3A1 mutation influences life expectancy.

“the nature of the specific COL3A1 mutation influences life expectancy.”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

May combine two claims.

This type of mutations produces a dominant negative effect on the protein, thereby inhibiting extracellular accumulation of mature COLLIII.

“This type of mutations produces a dominant negative effect on the protein, thereby inhibiting extracellular accumulation of mature COLLIII.”

• cites [8] LT Smith, 1997, Mutations in the COL3A1 gene result in the Ehlers-Danlos syndrome type IV and alterations in the size and distribution of the major collagen fibrils of the dermis, J Invest Dermatol sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

All experiments were repeated three times.

“All experiments were repeated three times.”

May combine two claims.

COL3A1 missense mutations altering glycine codons together with in-frame splice mutations in the triple helical domain lead to misfolding of COLLIII in the ER and retention of seven eighths of misfolded procollagen trimers in the cell with consequent enlargement of ER vesicles.

“COL3A1 missense mutations altering the glycine codons together with in-frame splice mutations in the triple helical domain account for the majority of disease-causing variants and lead to misfolding of COLLIII in the endoplasmic reticulum (ER) and retention of seven eighths of misfolded procollagen trimers in the cell with consequent enlargement of ER vesicles [8–10].”

• cites [8] LT Smith, 1997, Mutations in the COL3A1 gene result in the Ehlers-Danlos syndrome type IV and alterations in the size and distribution of the major collagen fibrils of the dermis, J Invest Dermatol sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Relative expression levels of a series of selected DEGs identified by array analysis were confirmed by quantitative real-time PCR (qPCR) using different RNA extractions obtained from skin fibroblast cultures of patients and healthy subjects.

“Relative expression levels of a series of selected DEGs identified by array analysis were confirmed by quantitative real-time PCR (qPCR) using different RNA extractions obtained from skin fibroblast cultures of patients and healthy subjects.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Biosynthesis of mature COLL fibrils is a multistep process spanning both intracellular and extracellular PTMs mediated by various enzymes and molecular chaperones.

“Biosynthesis of mature COLL fibrils is a multistep process spanning both intracellular and extracellular PTMs mediated by various enzymes and molecular chaperones that provide specific modifications such as hydroxylation of prolyl and lysyl residues, which are a prerequisite for correct protein folding [7].”

• cites [7] JF Bateman, 2009, Genetic diseases of connective tissues: cellular and extracellular effects of ECM mutations, Nat Genet sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Down-regulated LOXL3 expression in vEDS fibroblasts and the concomitant COLLIII- and ELN-ECM disorganization suggest that in dermal fibroblasts collagenogenesis and elastogenesis should be co-regulated.

“The down-regulated LOXL3 expression in vEDS fibroblasts and the concomitant COLLIII- and ELN-ECM disorganization suggest that in dermal fibroblasts collagenogenesis and elastogenesis should be co-regulated, according to Aziz and colleagues’ findings [38].”

• cites [38] J Aziz, 2016, Molecular Mechanisms of Stress-Responsive Changes in Collagen and Elastin Networks in Skin, Skin Pharmacol Physiol sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Our protein data indicated the disorganization of the core proteins of PGs perlecan, versican, and DCN, and their abnormal organization could contribute to the defective COLLIII fibril assembly in vEDS cells, as well as to the defective fibrillogenesis of other COLLs.

“Our protein data indicated the disorganization of the core proteins of PGs perlecan, versican, and DCN, which enhance COLL fibril formation [26], and their abnormal organization could contribute to the defective COLLIII fibril assembly in vEDS cells, as well as to the defective fibrillogenesis of other COLLs previously described in skin fibroblasts of different EDS types and in other connective”

• cites [16] N Zoppi, 2004, Human fibroblasts with mutations in COL5A1 and COL3A1 genes do not organize collagens and fibronectin in the extracellular matrix, down-regulate alpha2beta1 integrin, and recruit alphavbeta3 Instead of alpha5beta1 integrin, J Biol Chem sourceNot checked yet

  Not checked in the free run.

• cites [17] N Zoppi, 2008, FAK-independent alphavbeta3 integrin-EGFR complexes rescue from anoikis matrix-defective fibroblasts, Biochim Biophys Acta sourceNot checked yet

  Not checked in the free run.

• cites [19] N Zoppi, 2015, GLUT10 deficiency leads to oxidative stress and non-canonical αvβ3 integrin-mediated TGFβ signalling associated with extracellular matrix disarray in arterial tortuosity syndrome skin fibroblasts, Hum Mol Genet sourceNot checked yet

  Not checked in the free run.

• cites [33] M Baumann, 2012, Mutations in FKBP14 cause a variant of Ehlers-Danlos syndrome with progressive kyphoscoliosis, myopathy, and hearing loss, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [34] M Ritelli, 2014, Insights in the etiopathology of galactosyltransferase II (GalT-II) deficiency from transcriptome-wide expression profiling of skin fibroblasts of two sisters with compound heterozygosity for two novel B3GALT6 mutations, Mol Genet Metab RepNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Fibrillin microfibrils are ECM structures with essential functions in the development and organization of several tissues including blood vessels, and they can bind a number of other extracellular proteins, including latent TGFβ binding proteins, fibulins, and laminins.

“Fibrillin microfibrils are ECM structures with essential functions in the development and the organization of several tissues including blood vessels, and they can bind a number of other extracellular proteins, including latent TGFβ binding proteins (LTBPs), fibulins, and laminins [28].”

• cites [28] L Zilberberg, 2012, Specificity of latent TGF-β binding protein (LTBP) incorporation into matrix: role of fibrillins and fibronectin, J Cell Physiol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

The human COLLIII interactome revealed a nearly identical structural arrangement and functional domains as for COLLI, including cell interaction, fibrillogenesis and enzyme cleavage domains, intermolecular crosslink sites, and several MLBRs for different ECM components.

“More recently, the definition of the human COLLIII interactome revealed a nearly identical structural arrangement and functional domains as for COLLI, including cell interaction, fibrillogenesis and enzyme cleavage domains, intermolecular crosslink sites, and several MLBRs for different ECM components, such as other COLLs, PGs, and FN [24].”

• cites [24] JD Parkin, 2017, The collαgen III fibril has a "flexi-rod" structure of flexible sequences interspersed with rigid bioactive domains including two with hemostatic roles, PLoS One sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

The limited number of analyzed samples is a limitation of the study.

“We recognize the limited number of analyzed samples, as well as it is not obvious that other ECM producing cells, i.e., vascular smooth muscle cells and arterial fibroblasts, could show the same results in terms of gene expression profile.”

May combine two claims.

The strong retention of COLLIII in vEDS fibroblasts' cytoplasm suggests a role of FKBP22 in the biosynthesis and secretion mechanism of this COLL, and the reduction of this enzyme may represent a common trait of in vitro cultured fibroblasts derived from individuals with kEDS and vEDS.

“The strong retention of COLLIII in vEDS fibroblasts’ cytoplasm here reported suggests a role of FKBP22 in the biosynthesis and secretion mechanism of this COLL, and the reduction of this enzyme may represent a common trait of in vitro cultured fibroblasts derived from individuals with kEDS and vEDS.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Perlecan is a key component of the vascular ECM with endothelial barrier function, and it is a potent inhibitor of smooth muscle cell proliferation, thus contributing to maintain vascular homeostasis.

“It is also a key component of the vascular ECM with endothelial barrier function, and it is a potent inhibitor of smooth muscle cell proliferation, thus contributing to maintain vascular homeostasis [32].”

• cites [32] MS Lord, 2014, The role of vascular-derived perlecan in modulating cell adhesion, proliferation and growth factor signaling, Matrix Biol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Biallelic mutations in FKBP14 cause kyphoscoliotic EDS (kEDS-FKBP14) that is complicated by life-threatening cardiovascular complaints.

“Biallelic mutations in FKBP14 cause kyphoscoliotic EDS (kEDS-FKBP14) that is complicated by life-threatening cardiovascular complaints.”

• cites [33] M Baumann, 2012, Mutations in FKBP14 cause a variant of Ehlers-Danlos syndrome with progressive kyphoscoliosis, myopathy, and hearing loss, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [43] ML Murray, 2014, FKBP14-related Ehlers-Danlos syndrome: expansion of the phenotype to include vascular complications, Am J Med Genet A sourceNot checked yet

  Not checked in the free run.

• cites [44] C Dordoni, 2016, Further delineation of FKBP14-related Ehlers-Danlos syndrome: A patient with early vascular complications and non-progressive kyphoscoliosis, and literature review, Am J Med Genet A sourceNot checked yet

  Not checked in the free run.

• cites [45] C Giunta, 2017, A cohort of 17 patients with kyphoscoliotic Ehlers-Danlos syndrome caused by biallelic mutations in FKBP14: expansion of the clinical and mutational spectrum and description of the natural history, Genet MedNot checked yet

  Not checked in the free run.

May combine two claims.

In vEDS patients' skin fibroblasts, abnormal synthesis and deposition of COLLIII into the ECM together with disarray of FN and lack of COLLs- and FN-specific integrin receptors were previously demonstrated.

“In vEDS patients’ skin fibroblasts, we previously demonstrated an abnormal synthesis and deposition of COLLIII into the ECM together with the disarray of FN and the lack of COLLs- and FN-specific integrin receptors [16,17].”

• cites [16] N Zoppi, 2004, Human fibroblasts with mutations in COL5A1 and COL3A1 genes do not organize collagens and fibronectin in the extracellular matrix, down-regulate alpha2beta1 integrin, and recruit alphavbeta3 Instead of alpha5beta1 integrin, J Biol Chem sourceNot checked yet

  Not checked in the free run.

• cites [17] N Zoppi, 2008, FAK-independent alphavbeta3 integrin-EGFR complexes rescue from anoikis matrix-defective fibroblasts, Biochim Biophys Acta sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Hydroxylation of COLLs prolyl residues is essential for proper assembly and thermal stabilization of COLLs triple helices through intramolecular hydrogen bonding.

“Hydroxylation of COLLs prolyl residues is essential for proper assembly and thermal stabilization of COLLs triple helices through intramolecular hydrogen bonding [40], since under-hydroxylation results in abnormal assembly, and reduced thermal stability of newly synthesized procollagen chains [39].”

• cites [40] RA Berg, 1973, The thermal transition of a non-hydroxylated form of collagen. Evidence for a role for hydroxyproline in stabilizing the triple-helix of collagen, Biochem Biophys Res Commun sourceNot checked yet

  Not checked in the free run.

• cites [39] RA Gjaltema, 2017, Molecular insights into prolyl and lysyl hydroxylation of fibrillar collagens in health and disease, Crit Rev Biochem Mol Biol sourceNot checked yet

  Not checked in the free run.

May combine two claims.

The ER cisternae of FKBP22-deficient fibroblasts were dilated and filled of proteins.

“Moreover, the ER cisternae of FKBP22-deficient fibroblasts were dilated and filled of proteins [33].”

• cites [33] M Baumann, 2012, Mutations in FKBP14 cause a variant of Ehlers-Danlos syndrome with progressive kyphoscoliosis, myopathy, and hearing loss, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

ET-1 causes vasoconstriction mainly via ET-A, which represents the principle subtype in the vascular medial layer of large arteries.

“ET-1 causes vasoconstriction mainly via ET-A, which represents the principle subtype in the vascular medial layer of large arteries [36].”

• cites [36] AP Davenport, 2016, Endothelin, Pharmacol Rev sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

LOXL3 encodes a copper-dependent amine oxidase required for the formation of lysine-derived cross-links of COLLs and ELN fibrils.

“LOXL3 encodes a copper-dependent amine oxidase required for the formation of lysine-derived cross-links of COLLs and ELN fibrils [37].”

• cites [37] HM Kagan, 2003, Lysyl oxidase: properties, specificity, and biological roles inside and outside of the cell, J Cell Biochem sourceNot checked yet

  Not checked in the free run.

May combine two claims.

The present patients exhibited a full-blown expression of the disease with several major arterial events, in line with the limited genotype-phenotype correlation that associates this type of structural mutations with a more severe clinical phenotype.

“Besides, the present patients exhibited a full-blown expression of the disease with several major arterial events, in line with the limited genotype-phenotype correlation that associates this type of structural mutations with a more severe clinical phenotype [4,9–12].”

• cites [4] PH Byers, 2017, Diagnosis, natural history, and management in vascular Ehlers-Danlos syndrome, Am J Med Genet C Semin Med Genet sourceNot checked yet

  Not checked in the free run.

• cites [9] MG Pepin, 2014, Survival is affected by mutation type and molecular mechanism in vascular Ehlers–Danlos syndrome (EDS type IV), Genet Med sourceNot checked yet

  Not checked in the free run.

• cites [10] M Frank, 2015, The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome, Eur J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [11] U Schwarze, 2001, Haploinsufficiency for one COL3A1 allele of type III procollagen results in a phenotype similar to the vascular form of Ehlers-Danlos syndrome, Ehlers-Danlos syndrome type IV, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

• cites [12] DF Leistritz, 2011, COL3A1 haploinsufficiency results in a variety of Ehlers-Danlos syndrome type IV with delayed onset of complications and longer life expectancy, Genet Med sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

HSPG2 encodes perlecan, a PG present in the basement membranes, which binds to and cross-links many ECM components and cell-surface proteins.

“HSPG2 encodes perlecan, a PG present in the basement membranes, which binds to and cross-links many ECM components and cell-surface proteins, such as COLLI, COLLIV, laminin, nidogen, and FN [31].”

• cites [31] AJ Kvist, 2006, Chondroitin sulfate perlecan enhances collagen fibril formation. Implications for perlecan chondrodysplasias, J Biol Chem sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing), S4 (based).

It is not obvious that other ECM producing cells, i.e., vascular smooth muscle cells and arterial fibroblasts, could show the same results in terms of gene expression profile.

“as well as it is not obvious that other ECM producing cells, i.e., vascular smooth muscle cells and arterial fibroblasts, could show the same results in terms of gene expression profile.”

May combine two claims.

Since the patients harbored different structural COL3A1 mutations (glycine substitution and in-frame exon skipping), which correspond to the most frequent type of disease-causing variants, the present results should be representative of the majority of vEDS individuals.

“In addition, since our patients harbored different structural COL3A1 mutations, i.e., glycine substitution and in-frame exon skipping, which correspond to the most frequent type of the disease-causing variants, the present results should be representative of the majority of vEDS individuals.”

• AssumptionRests on an assumption: Glycine substitution and in-frame exon skipping correspond to the most frequent type of disease-causing COL3A1 variants.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

In FKBP22-deficient skin fibroblasts, a disturbed distribution and assembly of several ECM proteins has been described, suggesting that FKBP22 deficiency influences the proper deposition of different structural constituents of the ECM.

“In FKBP22-deficient skin fibroblasts a disturbed distribution and assembly of several ECM proteins, such as COLLI, COLLIII, COLLV, COLLVI, FN, tenascins, and thrombospondin has been described, suggesting that FKBP22 deficiency influences the proper deposition of different structural constituents of the ECM [33].”

• cites [33] M Baumann, 2012, Mutations in FKBP14 cause a variant of Ehlers-Danlos syndrome with progressive kyphoscoliosis, myopathy, and hearing loss, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

May combine two claims.

In vEDS skin fibroblasts, defective synthesis of COLLIII did not interfere with COLLV secretion and its extracellular organization, but altered the deposition and proper assembly into the ECM of COLLI, which was mainly retained in the cytoplasm.

“We previously showed that in vEDS skin fibroblasts the defective synthesis of COLLIII did not interfere with the COLLV secretion and its extracellular organization, but altered the deposition and proper assembly into the ECM of COLLI, which was mainly retained in the cytoplasm [16].”

• cites [16] N Zoppi, 2004, Human fibroblasts with mutations in COL5A1 and COL3A1 genes do not organize collagens and fibronectin in the extracellular matrix, down-regulate alpha2beta1 integrin, and recruit alphavbeta3 Instead of alpha5beta1 integrin, J Biol Chem sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Our protein data showed a generalized disassembly of all isotypes of fibrillin in vEDS fibroblasts' ECM, concomitant to the disorganization of EMILINs and ELN networks, providing an in vitro picture consistent with the pathological conditions of arterial walls reported in vEDS patients.

“Our protein data showed a generalized disassembly of all isotypes of fibrillin in vEDS fibroblasts’ ECM, concomitant to the disorganization of EMILINs and ELN networks, providing an in vitro picture consistent with the pathological conditions of arterial walls reported in vEDS patients.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

P4HA2 and P4HA3 encode two of the three isoforms of the α-subunit of collagen prolyl-4-hydroxylase, a tetrameric enzyme resident in the ER lumen that hydroxylates prolines on the procollagen peptide and serves as a chaperone for procollagen.

“P4HA2 and P4HA3 encode two of the three isoforms of the α-subunit of collagen prolyl-4-hydroxylase, a tetrameric enzyme resident in the ER lumen that hydroxylates prolines on the procollagen peptide and serves as a chaperone for procollagen [39].”

• cites [39] RA Gjaltema, 2017, Molecular insights into prolyl and lysyl hydroxylation of fibrillar collagens in health and disease, Crit Rev Biochem Mol Biol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

EDNRA encodes the endothelin receptor type A (ET-A) that interacts with receptor type B to mediate endothelin-1 signaling.

“EDNRA encodes the endothelin receptor type A (ET-A) that interacts with receptor type B to mediate endothelin-1 (ET-1) signaling, a potent endothelium-derived vasoconstrictor peptide that can activate vascular smooth muscle cells, inducing proliferation, and synthesis of ECM proteins [35].”

• cites [35] J Rodriguez-Vita, 2005, Endothelin-1, via ETA receptor and independently of transforming growth factor-beta, increases the connective tissue growth factor in vascular smooth muscle cells, Circ Res sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim. May combine two claims.

FKBP14 encodes FKBP22, a peptidyl-prolyl cis-trans isomerase localized in the ER lumen that interacts with different types of COLLs and catalyzes cis-trans-isomerization of peptide bonds, promoting COLLIII folding.

“FKBP14 encodes FKBP22, a peptidyl-prolyl cis-trans isomerase localized in the ER lumen [41] that interacts with different types of COLLs, i.e., COLLIII, COLLVI, and COLLX, and during COLLs triple helix formation catalyzes cis–trans-isomerization of peptide bonds, promoting in particular the COLLIII folding [39,42].”

• cites [41] I Raykhel, 2007, A molecular specificity code for the three mammalian KDEL receptors, J Cell Biol sourceNot checked yet

  Not checked in the free run.

• cites [39] RA Gjaltema, 2017, Molecular insights into prolyl and lysyl hydroxylation of fibrillar collagens in health and disease, Crit Rev Biochem Mol Biol sourceNot checked yet

  Not checked in the free run.

• cites [42] Y Ishikawa, 2014, A substrate preference for the rough endoplasmic reticulum resident protein FKBP22 during collagen biosynthesis, J Biol Chem sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Fibrillins interact closely with tropoelastin and integrins, and provide a scaffold for elastogenesis in elastic tissues.

“Fibrillins interact closely with tropoelastin and integrins, and provide a scaffold for elastogenesis in elastic tissues [26].”

• cites [26] J Halper, 2014, Basic components of connective tissues and extracellular matrix: elastin, fibrillin, fibulins, fibrinogen, fibronectin, laminin, tenascins and thrombospondins, Adv Exp Med Biol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Fibrillin-2 interacts directly with fibrillin-1, EMILIN-1, and LTBPs to regulate the bioavailability of TGFβ.

“Besides, fibrillin-2 interacts directly with fibrillin-1, EMILIN-1, and LTBPs to regulate the bioavailability of TGFβ, a growth factor and key regulator of connective tissue organization and blood vessels development and maintenance [27,29].”

• cites [27] MR Davis, 2012, Structure and function of the mammalian fibrillin gene family: Implications for human connective tissue diseases, Mol Genet Metabol sourceNot checked yet

  Not checked in the free run.

• cites [29] A Randell, 2017, Elastin microfibril interface-located protein 1, transforming growth factor beta, and implications on cardiovascular complications, J Am Soc Hypertens sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

ER stress is a mechanism involved in the pathogenesis of a range of systemic diseases including connective tissue disorders.

“increasing observations indicate ER stress as a mechanism involved in the pathogenesis of a range of systemic diseases including connective tissue disorders [56–59]”

• cites [56] RP Boot-Handford, 2010, The unfolded protein response and its relevance to connective tissue diseases, Cell Tissue Res sourceNot checked yet

  Not checked in the free run.

• cites [57] S De Palma, 2014, Muscle proteomics reveals novel insights into the pathophysiological mechanisms of collagen VI myopathies, J Proteome Res sourceNot checked yet

  Not checked in the free run.

• cites [58] M Pieri, 2014, Evidence for activation of the unfolded protein response in collagen IV nephropathies, J Am Soc Nephrol sourceNot checked yet

  Not checked in the free run.

• cites [59] A Hughes, 2017, Endoplasmic Reticulum Stress and Unfolded Protein Response in Cartilage Pathophysiology; Contributing Factors to Apoptosis and Osteoarthritis, Int J Mol Sci sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

The histone expression changes could reflect a diminished DNA synthesis.

“that could reflect a diminished DNA synthesis”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

PDIA4, PDIA5, and PDIA6 are members of an expanding protein family known as protein disulfide isomerases that may also act as effectors of protein translocation of terminally un/misfolded proteins to the cytosol for their proteasomal degradation.

“Moreover, PDIA4, PDIA5, and PDIA6 are members of an expanding proteins family known as protein disulfide isomerases that may also act as effectors of protein translocation of terminally un/misfolded proteins to the cytosol for their proteasomal degradation [50].”

• cites [50] AI Soares Moretti, 2017, Protein disulfide isomerases: Redox connections in and out of the endoplasmic reticulum, Arch Biochem Biophys sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

An anti-apoptotic role of FKBP22, favoring cell proliferation, by the over-expression of Bcl-2 and the silencing of caspases has been reported; conversely, shRNA-mediated FKBP14 inhibition induces cell cycle arrest and apoptosis.

“Recently, an anti-apoptotic role of FKBP22, favoring cell proliferation, by the over-expression of Bcl-2 and the silencing of caspases has been reported. Conversely, the shRNA-mediated FKBP14 inhibition induces cell cycle arrest and apoptosis [69]”

• cites [69] M Lu, 2016, RNAi-Mediated Downregulation of FKBP14 Suppresses the Growth of Human Ovarian Cancer Cells, Oncol Res sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Lack of classical UPR activation (e.g., elevated BiP levels) was previously reported in skin fibroblasts of Osteogenesis imperfecta patients with mutations in triple helical domain of COL1A1, in embryonic fibroblasts from a Marfan syndrome mouse model, and in patients' fibroblasts with different types of mucopolysaccharidoses.

“the lack of a classical UPR activation (e.g., elevated BiP levels) was previously reported in skin fibroblasts of Osteogenesis imperfecta patients with mutations in triple helical domain of COL1A1 [60], as well as in embryonic fibroblasts from a Marfan syndrome mouse model [61] and in patients’ fibroblasts with different types of mucopolysaccharidoses [62]”

• cites [60] SD Chessler, 1993, BiP binds type I procollagen proα chains with mutations in the carboxyl-terminal propeptides synthesized by cells from patients with osteogenesis imperfecta, J Biol Chem sourceNot checked yet

  Not checked in the free run.

• cites [61] T Meirelles, 2016, Fibrillin-1 mgΔ(lpn) Marfan syndrome mutation associates with preserved proteostasis and bypass of a protein disulfide isomerase-dependent quality checkpoint, Int J Biochem Cell Biol sourceNot checked yet

  Not checked in the free run.

• cites [62] GR Villani, 2012, Unfolded protein response is not activated in the mucopolysaccharidoses but protein disulfide isomerase 5 is deregulated, J Inherit Metab Dis sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Histones enter nucleosome formation and proper DNA wrapping after the S phase of cell cycle.

“histones enter in nucleosome formation and proper DNA wrapping after the S phase of cell cycle [65]”

• cites [65] GE Zentner, 2013, Regulation of nucleosome dynamics by histone modifications, Nat Struct Mol Biol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Kinesin family members are a class of motor proteins involved in cell movements and intracellular vesicles' trafficking from ER, and in chromosome and centrosome positioning during mitosis.

“a class of motor proteins involved in cell movements and intracellular vesicles’ trafficking from ER, and in chromosome and centrosome positioning during mitosis [66]”

• cites [66] KJ Verhey, 2011, Kinesin Assembly and Movement in Cells, Annual Review of Biophysics sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Procollagen misfolding caused by mutations within the triple helix domain likely does not activate UPR, but rather leads to the accumulation of insoluble procollagen aggregates that could be eliminated by autophagy.

“it is likely that procollagen misfolding caused by mutations within the triple helix domain does not activate UPR [60,63], but rather leads to the accumulation of insoluble procollagen aggregates that could be eliminated by autophagy [64]”

• cites [60] SD Chessler, 1993, BiP binds type I procollagen proα chains with mutations in the carboxyl-terminal propeptides synthesized by cells from patients with osteogenesis imperfecta, J Biol Chem sourceNot checked yet

  Not checked in the free run.

• cites [63] A Forlino, 2007, Differential expression of both extracellular and intracellular proteins is involved in the lethal or nonlethal phenotypic variation of BrtllV, a murine model for osteogenesis imperfecta, Proteomics sourceNot checked yet

  Not checked in the free run.

• cites [64] Y Ishida, 2009, Autophagic elimination of misfolded procollagen aggregates in the endoplasmic reticulum as a means of cell protection, Mol Cell Biol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

snoRNAs are a class of ncRNAs involved in the ribosomal RNA maturation and biogenesis.

“a class of ncRNAs involved in the ribosomal RNA maturation and biogenesis [70]”

• cites [70] W Filipowicz, 2002, Review: Biogenesis of small nucleolar ribonucleoproteins, Curr Opin Cell Biol sourceNot checked yet

  Not checked in the free run.

May combine two claims.

In patients' fibroblasts, the DNA damage response and the cyclins/CDKs/CDK inhibitors machine seem to be perturbed.

“Also, in patients’ fibroblasts the DNA damage response and the cyclins/CDKs/CDK inhibitors machine seem to be perturbed.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Structural mutations in genes encoding for different COLLs, i.e., COLLI, COLLIII, and COLLV, affect not only the assembly but also the secretion pathways of other ECM components in EDS fibroblasts.

“according to the notion that structural mutations in genes encoding for different COLLs, i.e., COLLI, COLLIII, and COLLV, affect not only the assembly but also the secretion pathways of other ECM components in EDS fibroblasts [16,17,33].”

• cites [16] N Zoppi, 2004, Human fibroblasts with mutations in COL5A1 and COL3A1 genes do not organize collagens and fibronectin in the extracellular matrix, down-regulate alpha2beta1 integrin, and recruit alphavbeta3 Instead of alpha5beta1 integrin, J Biol Chem sourceNot checked yet

  Not checked in the free run.

• cites [17] N Zoppi, 2008, FAK-independent alphavbeta3 integrin-EGFR complexes rescue from anoikis matrix-defective fibroblasts, Biochim Biophys Acta sourceNot checked yet

  Not checked in the free run.

• cites [33] M Baumann, 2012, Mutations in FKBP14 cause a variant of Ehlers-Danlos syndrome with progressive kyphoscoliosis, myopathy, and hearing loss, Am J Hum Genet sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Cell cycle progression in eukaryotic cells is strictly regulated by both integrin-mediated adhesion to the ECM and by binding of growth factors to their receptors.

“It is well known that cell cycle progression in eukaryotic cells is strictly regulated by both integrin-mediated adhesion to the ECM and by binding of growth factors to their receptors [68]”

• cites [68] MA Schwartz, 2001, Integrins and cell proliferation: regulation of cyclin-dependent kinases via cytoplasmic signaling pathways, J Cell Sci sourceNot checked yet

  Not checked in the free run.

May combine two claims.

vEDS fibroblasts undergo an ECM-deficiency-mediated pre-apoptotic state, in which caspases' activity is up-regulated according to the down-expression of Bcl-2.

“they undergo an ECM-deficiency-mediated pre-apoptotic state, in which caspases’ activity is up-regulated according to the down-expression of Bcl-2 [17]”

• cites [17] N Zoppi, 2008, FAK-independent alphavbeta3 integrin-EGFR complexes rescue from anoikis matrix-defective fibroblasts, Biochim Biophys Acta sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

This cell cycle phases' perturbation could be a consequence of accumulated DNA mismatches, probably not repaired by not-functioning mismatch repair mechanism.

“This cell cycle phases’ perturbation could be a consequence of accumulated DNA mismatches, probably not repaired by not-functioning mismatch repair mechanism.”

May combine two claims.

COL3A1 mismatches can be responsible for misfolded COLLIII trimers, which are not processed by proteasome and accumulate in the ER enlarged cisternae blocking vEDS cells' trafficking.

“COL3A1 mismatches can be responsible of misfolded COLLIII trimers, which are not processed by proteasome and accumulate in the ER enlarged cisternae blocking vEDS cells’ trafficking.”

May combine two claims.

Enlargement of ER vesicles entails an accumulation of misfolded proteins, including COLLIII, and ensuing impairment of protein folding quality control.

“which should entail an accumulation of misfolded proteins, including COLLIII, and ensuing impairment of protein folding quality control”

Probably not a substantial claim: may not pass S3 (load-bearing).

Various types of ncRNAs including snoRNAs can also participate in the regulation of gene expression, cell cycle, DNA repair, genome integrity, as well as in human diseases.

“Accumulating evidence suggested that various types of ncRNAs including snoRNAs can also participate in the regulation of gene expression, cell cycle, DNA repair, genome integrity, as well as in human diseases [71,72]”

• cites [71] G Wan, 2014, Noncoding RNAs in DNA repair and genome integrity, Antioxid Redox Signal sourceNot checked yet

  Not checked in the free run.

• cites [72] GA Stepanov, 2015, Regulatory role of small nucleolar RNAs in human diseases, Biomed Res Int sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

Disturbance of ER redox state, as a consequence of oxidative stress, alteration of calcium homeostasis, or dysregulated chaperones activity, may result in un/misfolded, or aggregated proteins that will be targeted to degradation pathways or accumulate in cells.

“Disturbance of ER redox state, as a consequence of oxidative stress, alteration of calcium homeostasis, or dysregulated chaperones activity, may result in un/misfolded, or aggregated proteins that will be targeted to degradation pathways or accumulate in cells [47].”

• cites [47] E Ullman, 2008, Autophagy promotes necrosis in apoptosis-deficient cells in response to ER stress, Cell Death Differ sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

UPR is an adaptive pathway to restore protein folding equilibrium.

“an adaptive pathway to restore protein folding equilibrium [51]”

• cites [51] T Anelli, 2008, Protein quality control in the early secretory pathway, EMBO J sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The ER not only provides mechanisms to facilitate proper folding of newly synthesized secretory and membrane proteins, but also harbors molecular machineries that eliminate proteins that fail to fold or assemble correctly.

“ER not only provides mechanisms to facilitate proper folding of newly synthesized secretory and membrane proteins, but also harbors molecular machineries that eliminate proteins that fail to fold or assemble correctly [46].”

• cites [46] RS Hegde, 2010, Quality and quantity control at the endoplasmic reticulum, Curr Opin Cell Biol sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

These processes (DNA repair and cell cycle) are commonly damaged in primary human fibroblast strains undergoing a cellular aging process as a consequence of finite proliferative capacity or oxidative stress.

“These processes are commonly damaged in primary human fibroblast strains undergoing a cellular aging process as a consequence of finite proliferative capacity or oxidative stress [67]”

• cites [67] S Marthandan, 2016, Conserved genes and pathways in primary human fibroblast strains undergoing replicative and radiation induced senescence, Biol Res sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The proteasome complex participates in the maintenance of protein homeostasis, and its dysfunction contributes to the accumulation of protein aggregates.

“a proteolytic machinery that participates in the maintenance of protein homeostasis and its dysfunction contributes to the accumulation of protein aggregates [51]”

• cites [51] T Anelli, 2008, Protein quality control in the early secretory pathway, EMBO J sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Previous study demonstrated in vEDS fibroblasts a decrease of Bcl-2 protein expression and activation of caspase enzymes.

“our previous study demonstrated in vEDS fibroblasts the decrease of Bcl-2 protein expression and the activation of caspase enzymes”

• cites [17] N Zoppi, 2008, FAK-independent alphavbeta3 integrin-EGFR complexes rescue from anoikis matrix-defective fibroblasts, Biochim Biophys Acta sourceNot checked yet

  Not checked in the free run.