Presentation number: MG 23
TWO RARE CASES OF CEREBROTENDINOUS XANTHOMATOSIS IN THE SAME FAMILY CAUSED BY AN INTRONIC MUTATION IN CYP27A1
Petar Brlek1, Zrinka Djukić Koroljević1, Ivica Grgurević1, Jelena Bošnjak1, Andrea Skelin1, Tomislav Pavlović1, Igor Borić1,2,3, Ivo Barić4, Dragan Primorac1,2,3,5,6,7,8,9,10
1St. Catherine Specialty Hospital, Zagreb, Croatia, 2Medical School, University of Rijeka, Rijeka, Croatia, 3Medical School, University of Mostar, Mostar, Bosnia and Herzegovina, 4Department of Paediatrics, University Hospital Center Zagreb and University of Zagreb School of Medicine, Zagreb, Croatia, 5Medical School, University of Split, Split, Croatia , 6Faculty of Dental Medicine and Health, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia, 7Department of Biochemistry & Molecular Biology, The Pennsylvania State University, State College, PA, USA, 8The Henry C. Lee College of Criminal Justice and Forensic Sciences, University of New Haven, West Haven, CT, USA, 9Medical School REGIOMED, Coburg, Germany, 10National Forensic Sciences University, Gujarat, India
Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive lipid storage disease associated with abnormally high blood cholestanol levels. Sterol 27-hydroxylase enzyme deficiency leads to abnormal metabolization of cholesterol and diverts the metabolic pathway in the direction of reduced production of chenodeoxycholic acid and an increased formation of 25-hydroxylated bile alcohols and cholestanol. CTX patients commonly have infantile-onset diarrhea, childhood-onset cataract, adult-onset progressive neurologic symptoms, and adolescent to young adult-onset tendon xanthomas. Cholestanol accumulates in the brain, tendons, eyes, and blood vessels, causing a range of clinical manifestations, including ataxia, epilepsy, dementia, and even parkinsonism. We present a case of two brothers aged 34 and 38 with a similar clinical picture that includes ataxia, cognitive deficit, congenital cataract, and diarrhea since birth. The older brother also experienced grand mal seizures. Parents are healthy and are not in consanguinity. Moreover, other family members are without neuromuscular diseases and known heredity in the family. Patients came to the St. Catherine Specialty Hospital to perform genetic testing and counseling due to a suspicion of a genetic disorder. Next-generation sequencing of 444 genes associated with various neurological and neuromuscular disorders with clinical examination, radiological and biochemical analysis established a diagnosis of autosomal recessive cerebrotendinous xanthomatosis caused by a homozygous form of variant c.1184+1G>A in both brothers. This mutation affects a donor splice site in intron 6 of the CYP27A1 gene. Studies have shown that disruption of this splice site results in the skipping of 89 nucleotides of exon six and introduces a premature termination codon which results in mRNA nonsense-mediated decay. However, replacement therapy with chenodeoxycholic acid may prevent clinical deterioration. Early treatment in symptomatic patients was shown to stop progression and, in some cases, reduction of pre-existing neurological deficits. Patients were referred to systematic clinical monitoring and treatment under the supervision of the clinical medical team at St. Catherine Specialty Hospital.
Key words: sterol 27-hydroxylase, cholestanol, metabolic disease, CYP27A1, chenodeoxycholic acid
Presentation number: MG 24
QUANTITATIVE ANALYSIS OF MIRNA IN SUDDEN CARDIAC DEATH TISSUE AND BLOOD SAMPLES
Luise Mildeberger, Julia Bueto, Verena Wilmes, Stefanie Scheiper-Welling, Marcel A. Verhoff, Silke Kauferstein
University Hospital Frankfurt, Institute of Legal Medicine, Frankfurt am Main, Germany
Cardiovascular diseases are the leading cause of death worldwide. Currently, microRNAs (miRNAs) are very promising biomarkers in various cardiovascular diseases. MiRNAs are short non-coding RNAs, which regulate a vast variety of biological processes. They bind to their target mRNA at the 3’UTR and can either inhibit translation or initiate their degradation. The focus of the current study was to investigate the expression of different miRNAs in heart tissue and whole blood samples from sudden cardiac death (SCD) in comparison to control hearts. The expression of different miRNAs in heart tissue and whole blood of SCD and control cases was assessed via q-RT-PCR in accordance with the MIQE guidelines. SCD group was divided in cases with structural changes (myocardial infarction, MI) and cases without visible changes. Statistical evaluation of significance and receiver operating characteristic (ROC) analysis was performed via R Studio. An upregulation of miR-1, miR-26a and miR-133a in tissue of SCD samples without structural changes compared to MI and controls were found. Furthermore, miR-1 and miR-133a were upregulated in MI whole blood. Receiver operating characteristics show a better diagnostic accuracy of miR-1 and miR-133a in whole blood than in tissue. MiR-1 and miR-133a were among the first miRNAs to be discovered in mammals, but their physiological purpose still remains unclear. The results of this study reveal that whole blood is better suited for quantitative expression analysis of these miRNAs. Especially the muscle specific miRNAs (MyomiRs) miR-1 and miR-133a showed expression differences in tissue and whole blood of MI and SCD. The higher expressions of these miRNAs in whole blood after MI may point to necrosis of the heart tissue and therefore could be a useful degradation marker for acute MI. There are still challenges that need to be addressed to establish these new biomarkers. Nevertheless, this study may be the basis for further clinical and basic research in the field of blood-based miRNA-profiling in sudden cardiac death.
Key words: MicroRNA, biomarkers, sudden cardiac death, myocardial infarction
Presentation number: MG 25
TREATMENT OF PULMONARY SARCOIDOSIS USING ALLOGENIC BONE MARROW-DERIVED MESENCHYMAL STEM CELL THERAPY IS SAFE: A CASE REPORT
Eduard Pavelic1, Vid Matisic1, Vilim Molnar1, Petar Brlek1, Tomislav Pavlovic1, Marko Strbad2,3, Lenart Girandon2, Dragan Primorac1,4,5,6,7,8,9,10,11
1St Catherine Specialty Hospital, Department of Orthopedics, Zagreb, 2Educell Ltd, Trzin, Slovenia, 3Biobanka Ltd, Trzin, Slovenia, 4University of Split, School of Medicine, Split, Croatia, 5University of Split, University Department of Forensic Sciences, Split, Croatia, 6University of Osijek, Faculty of Medicine, Osijek, Croatia, 7University of Osijek, Faculty of Dental Medicine and Health, Osijek, Croatia, 8University of Rijeka, School of Medicine, Rijeka, Croatia, 9University of New Haven, Henry C. Lee College of Criminal Justice and Forensic Sciences, West Haven, CT, USA, 10Medical School REGIOMED, Coburg, Germany, 11The National Forensic Sciences University, Gandhinagar, Gujarat, India
In the 21rst century mesenchymal stem cells (MSC) are used from a variety of sources whether adipose derived, placental or bone marrow. These mesenchymal stem cells have a proven potent immunomodulator effect both in vitro and in vivo. The growing field of personalized medicine allowed for the introduction of stem cell therapy in the treatment of systemic and localized diseases. We report a case of a 67-year-old male patient, first diagnosed with sarcoidosis in 2005, that responded well clinically and biologically to ImmunoARTTM (developed by Educell Ltd., member of Medical Biobank Swiss Institute SA (MBSI)) allogenic, HLA-incompatible and non-related bone marrow-derived MSCs in a dose of 106/kg. The patient presented to St. Catherine Specialty Hospital in 2021 with an exacerbation of respiratory symptoms. After a clinical and radiological examination with laboratory workup, radiological findings were consistent with pulmonary sarcoidosis, while laboratory work revealed increased leucocytes at 14.2 g/L, CRP at 51.2 mg/L and lymphocytes at 4.25 g/L. The patient was then administered with intravenous application of MSCs on three occasions in the out-patient clinic. MSC doses were prepared from a young, healthy donor who agreed to donate bone marrow for allogeneic treatment and who was negative for viral markers (HBs Ag, HBc Ab, HCV Ab, HIV 1-2 Ab, TPHA, HBV NAT, HCV NAT, HIV NAT) according to EU legislation. Cells were prepared in a controlled and verified laboratory for “Hospital exemption” cell preparation in the cleanroom facility in safety cabinet class A and expressed CD105, CD 73 and CD 90 but lacked the expression of CD45 and CD34. Over the course of MSC therapy, the patient showed clinical and biological with a decrease in inflammatory parameters. Laboratory values were assessed at days 2, 5 and 7. On day 7, leucocytes were 11.2 g/L, lymphocytes 4.0 g/L and CRP 5.1 mg/L. In the short follow-up period the patient felt subjectively better, without any side effects to the MSC therapy. Unfortunately, the patient dropped out from follow up, therefore the prolonged effects of this therapy were not able to be assessed. Therefore, systemic MSC therapy presents an opportunity for treatment of sarcoidosis that needs to be further researched.
Key words: Mesenchymal stem cells, allogenic MSC, sarcoidosis