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Screening of a HUVEC cDNA library with transplant-associated coronary artery disease sera identifies RPL7 as a
A T Linke1, B Marchant, P Marsh
1National Heart and Lung Institute, Imperial College School of Medicine, Harefield Hospital, Middlesex, UK.
Insights
Researchers identified specific genes associated with transplant-associated coronary artery disease (TxCAD). Ribosomal protein L7 showed the highest prevalence in patients with TxCAD, suggesting its potential role in the disease.
Area of Science:
- Cardiovascular Biology
- Transplantation Immunology
- Molecular Biology
Background:
- Transplant-associated coronary artery disease (TxCAD) is a significant complication following cardiac transplantation.
- Identifying specific molecular markers for TxCAD is crucial for early diagnosis and management.
Purpose of the Study:
- To screen a HUVEC cDNA library using sera from TxCAD patients to identify potential disease-associated genes.
- To determine the prevalence and association of identified genes with TxCAD.
Main Methods:
- Screening of a HUVEC cDNA library with patient sera.
- DNA sequence analysis of positive clones.
- Expression of selected genes as recombinant fusion proteins.
- ELISA screening of a wider patient cohort.
Main Results:
- Six positive clones were isolated, including lysyl tRNA synthetase, ribosomal protein L7, ribosomal protein L9, beta transducin, and TANK.
- Ribosomal protein L7 demonstrated the highest prevalence (55.6%) in TxCAD patient sera compared to non-CAD controls (10%).
Conclusions:
- Ribosomal protein L7 is a potential biomarker for transplant-associated coronary artery disease.
- Further investigation into the role of identified genes in TxCAD pathogenesis is warranted.
Abstract:
A HUVEC cDNA library was screened with sera from two patients who had developed transplant-associated coronary artery disease (TxCAD) following cardiac transplantation. A total of six positive clones were isolated from a primary screen of 40 000 genes. Subsequent DNA sequence analysis identified these to be lysyl tRNA synthetase, ribosomal protein L7, ribosomal protein L9, beta transducin and TANK. Another gene whose product could not be identified showed homology to a human cDNA clone (DKFZp566M063) derived from fetal kidney. Full-length constructs of selected genes were expressed as his-tag recombinant fusion proteins and used to screen a wider patient base by ELISA to determine prevalence and association with TxCAD. Of these ribosomal protein L7 showed the highest prevalence (55.6%) with TxCAD sera compared to 10% non-CAD.