Naslov
RNA-Seq analysis of fibroblasts from patient with S- adenosylhomocysteine hydrolase deficiency

Autori
Šimunić, Ena ; Rokić, Filip ; Vugrek, Oliver

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
5. simpozij studenata doktorskih studija PMF-a : knjiga sažetaka = 5th PhD Student Symposium 2021 : book of abstracts / Barišić, Dajana - Zagreb, 2021, 247-247

ISBN
978-953-607690-1

Skup
5. Simpozij studenata doktorskih studija PMF-a = 5th PhD Student Symposium 2021

Mjesto i datum
Zagreb, Hrvatska, 24.04.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
RNA-seq ; AHCY

Sažetak
S-adenosylhomocystein hydrolase (AHCY) is an enzyme that catalyzes hydrolysis of S-adenosyl- Lhomocysteine (SAH) to adenosine and homocysteine. [1] Since SAH is the product and competitive inhibitor of all S-adenosyl methionine-dependent transmethylation reactions, its removal is crucial for maintaining cells’ methylation potential and normal organism function. [2] Reduced AHCY activity leads to inhibition of S-adenosyl methionine- dependent transmethylation reactions which leads to severe metabolic disorders and pathological conditions in humans. This clinical presentation is also characterized with increased concentrations of SAH, S-adenosyl methionine (SAM) and methionine in plasma. This severe and hereditary metabolic disease caused by insufficient AHCY activity was described for the first time in Croatia in 2004. [3] So far, several mutations that lead to reduced AHCY activity have been found (R49C, R49H, G71S, D86G, A89V, Y143C, Y328D and W112Ter). To gain insight into possible consequences on the patient’s transcriptome due to insufficient AHCY activity caused by mutations Y328D and Y143C, RNA extracted from patient’s dermal fibroblasts was sequenced (RNAseq.). This was done on Illumina NextSeq System, using three biological replicas. RNA extracted from healthy fibroblasts was used as control (WT). The obtained raw data was processed using commercially available applications RNA-Seq alignment (Illumina, Inc.), DeSeq 2 (BaseSpace Labs.) and iPathwayGuide (Advaita Bio). RNA-Seq alignment was used to align obtained sequences which were then analyzed with DeSeq2 to obtain a list of differentially expressed genes. 50 differentially expressed genes with highest absolute log2-fold change value (out of 1692 observed, with p<0, 05) were analyzed using iPathwayGuide. Cell pathways that can be assumed to be most altered in patient’s fibroblasts are PI3K-Akt signaling pathway, focal adhesion and ECM-receptor interaction. Based on this results it can be concluded that cell cycle regulation and synthesis of collagens, laminins and integrins is disturbed in patient’s fibroblasts. Disturbed regulation of key extracellular proteins might lead to infringed adhesion and cell-extracellular matrix interaction.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA