Cell Line and DNA Biobank From Patients Affected by Genetic Diseases

Filocamo, Mirella; Mazzotti, Raffaella; Corsolini, Fabio; Stroppiano, Marina; Stroppiana, Giorgia; Grossi, Serena; Lualdi, Susanna; Tappino, Barbara; Lanza, Federica; Galotto, Sara; Biancheri, Roberta

doi:10.5334/ojb.ab

Cited by 32 publications

(14 citation statements)

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“…NPC patients were diagnosed by the test with filipin and confirmed by mutation analysis in the NPC1 and NPC2 genes. Skin fibroblasts lines from GD and controls originated from the collection of Institute of Psychiatry and Neurology, Department of Genetics (Warsaw, Poland), while the NPC patients’ cell lines originated from the ‘Cell line and DNA Biobank from patients affected by Genetic Diseases’ located at G. Gaslini Institute (Genoa, Italy) 49 and member of the Telethon Network of Genetic Biobanks (http://biobanknetwork.telethon.it/).…”

Section: Methodsmentioning

confidence: 99%

Gene expression profile in patients with Gaucher disease indicates activation of inflammatory processes

Ługowska

Hetmańczyk-Sawicka

Iwanicka-Nowicka

et al. 2019

Sci Rep

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Gaucher disease (GD) is a rare inherited metabolic disease caused by pathogenic variants in the GBA1 gene. So far, the pathomechanism of GD was investigated mainly in animal models. In order to delineate the molecular changes in GD cells we analysed gene expression profile in cultured skin fibroblasts from GD patients, control individuals and, additionally, patients with Niemann-Pick type C disease (NPC). We used expression microarrays with subsequent validation by qRT-PCR method. In the comparison GD patients vs. controls, the most pronounced relative fold change (rFC) in expression was observed for genes IL13RA2 and IFI6 (up-regulated) and ATOH8 and CRISPLD2 (down-regulated). Products of up-regulated and down-regulated genes were both enriched in genes associated with immune response. In addition, products of down-regulated genes were associated with cell-to-cell and cell-to-matrix interactions, matrix remodelling, PI3K-Akt signalling pathway and a neuronal survival pathway. Up-regulation of PLAU , IFIT1 , TMEM158 and down-regulation of ATOH8 and ISLR distinguished GD patients from both NPC patients and healthy controls. Our results emphasize the inflammatory character of changes occurring in human GD cells indicating that further studies on novel therapeutics for GD should consider anti-inflammatory agents.

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Section: Methodsmentioning

confidence: 99%

Gene expression profile in patients with Gaucher disease indicates activation of inflammatory processes

Ługowska

Hetmańczyk-Sawicka

Iwanicka-Nowicka

et al. 2019

Sci Rep

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“…ALG3-CDG (α-1,3-mannosyltransferase deficiency), PGAP2-CDG (post-GPI attachment to proteins 2 deficiency), ALG8-CDG (α-1,3-glucosyltransferase 2 deficiency), GMPPB-CDG (GDP-mannose pyrophosphorylase B) and control (CTR) fibroblasts from age-matched volunteers were provided by the Neuroscience Department of the Meyer Children’s Hospital (Florence, Italy). PMM2-CDG (phosphomannomutase-2 deficiency) fibroblasts were obtained from the Giannina Gaslini Institute-Telethon Network of Genetic Biobanks (Genoa, Italy) [ 30 ]. Clinical features of these patients have been previously published in part [ 31 ] and are described in the Supplementary Table 1.…”

Section: Methodsmentioning

confidence: 99%

Defective IGF-1 prohormone N-glycosylation and reduced IGF-1 receptor signaling activation in congenital disorders of glycosylation

Patria

Annibalini

Morrone

et al. 2022

Cell. Mol. Life Sci.

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The insulin-like growth factor-1 (IGF-1) signaling pathway is crucial for the regulation of growth and development. The correct processing of the IGF-1Ea prohormone (proIGF-1Ea) and the IGF-1 receptor (IGF-1R) peptide precursor requires proper N-glycosylation. Deficiencies of N-linked glycosylation lead to a clinically heterogeneous group of inherited diseases called Congenital Disorders of Glycosylation (CDG). The impact of N-glycosylation defects on IGF-1/IGF-1R signaling components is largely unknown. In this study, using dermal fibroblasts from patients with different CDG [PMM2-CDG (n = 7); ALG3-CDG (n = 2); ALG8-CDG (n = 1); GMPPB-CDG (n = 1)], we analyzed the glycosylation pattern of the proIGF-1Ea, IGF-1 secretion efficiency and IGF-1R signaling activity. ALG3-CDG, ALG8-CDG, GMPPB-CDG and some PMM2-CDG fibroblasts showed hypoglycosylation of the proIGF-1Ea and lower IGF-1 secretion when compared with control (CTR). Lower IGF-1 serum concentration was observed in ALG3-CDG, ALG8-CDG and in some patients with PMM2-CDG, supporting our in vitro data. Furthermore, reduced IGF-1R expression level was observed in ALG3-CDG, ALG8-CDG and in some PMM2-CDG fibroblasts. IGF-1-induced IGF-1R activation was lower in most PMM2-CDG fibroblasts and was associated with decreased ERK1/2 phosphorylation as compared to CTR. In general, CDG fibroblasts showed a slight upregulation of Endoplasmic Reticulum (ER) stress genes compared with CTR, uncovering mild ER stress in CDG cells. ER-stress-related gene expression negatively correlated with fibroblasts IGF-1 secretion. This study provides new evidence of a direct link between N-glycosylation defects found in CDG and the impairment of IGF-1/IGF-1R signaling components. Further studies are warranted to determine the clinical consequences of reduced systemic IGF-1 availability and local activity in patients with CDG.

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“…Primary Human Dermal Fibroblast (NDHF Promo Cell #FB60C12350) supplied by Carlo Erba were used as the control cell line and compared with primary dermal fibroblast from a young Marinesco-Sjögren syndrome patient, supplied by Telethon Network of Genetic Biobanks-TNGB [36]. Cells were cultured in Dulbecco's modified Eagle's medium + GlutaMAX (GIBCO, 61965-026), supplemented with 10% of Fetal Bovine Serum (FBS) and 1% penicillin/streptomycin PenStrep (GIBCO, 15070-063).…”

Section: Cell Culturementioning

confidence: 99%

Proteomic Analysis of Marinesco–Sjogren Syndrome Fibroblasts Indicates Pro-Survival Metabolic Adaptation to SIL1 Loss

Potenza

Cufaro

Biase

et al. 2021

IJMS

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Marinesco–Sjogren syndrome (MSS) is a rare multisystem pediatric disorder, caused by loss-of-function mutations in the gene encoding the endoplasmic reticulum cochaperone SIL1. SIL1 acts as a nucleotide exchange factor for BiP, which plays a central role in secretory protein folding. SIL1 mutant cells have reduced BiP-assisted protein folding, cannot fulfil their protein needs, and experience chronic activation of the unfolded protein response (UPR). Maladaptive UPR may explain the cerebellar and skeletal muscle degeneration responsible for the ataxia and muscle weakness typical of MSS. However, the cause of other more variable, clinical manifestations, such as mild to severe mental retardation, hypogonadism, short stature, and skeletal deformities, is less clear. To gain insights into the pathogenic mechanisms and/or adaptive responses to SIL1 loss, we carried out cell biological and proteomic investigations in skin fibroblasts derived from a young patient carrying the SIL1 R111X mutation. Despite fibroblasts not being overtly affected in MSS, we found morphological and biochemical changes indicative of UPR activation and altered cell metabolism. All the cell machineries involved in RNA splicing and translation were strongly downregulated, while protein degradation via lysosome-based structures was boosted, consistent with an attempt of the cell to reduce the workload of the endoplasmic reticulum and dispose of misfolded proteins. Cell metabolism was extensively affected as we observed a reduction in lipid synthesis, an increase in beta oxidation, and an enhancement of the tricarboxylic acid cycle, with upregulation of eight of its enzymes. Finally, the catabolic pathways of various amino acids, including valine, leucine, isoleucine, tryptophan, lysine, aspartate, and phenylalanine, were enhanced, while the biosynthetic pathways of arginine, serine, glycine, and cysteine were reduced. These results indicate that, in addition to UPR activation and increased protein degradation, MSS fibroblasts have profound metabolic alterations, which may help them cope with the absence of SIL1.

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Cell Line and DNA Biobank From Patients Affected by Genetic Diseases

Abstract: Biological samples and associated data.

Cited by 32 publications

References 1 publication

Gene expression profile in patients with Gaucher disease indicates activation of inflammatory processes

Gene expression profile in patients with Gaucher disease indicates activation of inflammatory processes

Defective IGF-1 prohormone N-glycosylation and reduced IGF-1 receptor signaling activation in congenital disorders of glycosylation

Proteomic Analysis of Marinesco–Sjogren Syndrome Fibroblasts Indicates Pro-Survival Metabolic Adaptation to SIL1 Loss

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