Volume 47, Issue 1, January 2024, Pages 24-31Author links open overlay panel, , , , , , AbstractBackground

MicroRNAs (miRNAs) are a group of small non-coding RNAs that bind to the target mRNA and regulate gene expression. Recently circulating microRNAs were investigated as markers of diseases and therapeutic targets. Although various studies analyze the miRNA expression in liver disease, these studies on PFIC are few. Progressive familial intrahepatic cholestasis (PFIC) is a rare liver disease with autosomal recessive inheritance. Most children with PFIC progress to cirrhosis and liver failure and consequently need to have a liver transplant. The aim of this study is the investigation of the miR-19b and miR-let7b expression levels in Iranian PFIC children.

Methods

25 PFIC patients, 25 healthy children and 25 Biliary Atresia patients were considered as case and two control groups respectively. Blood samples were obtained and Liver function tests (LFTs) were measured. After RNA extraction and cDNA synthesis, quantitative PCR was performed using specific primers for miR-19b and miR-let7b. The U6 gene is used as an internal control.

Results

qPCR on PFIC patients’ samples demonstrated that the miR-19b and the miR-let7b expression were significantly decreased in patients compared to the control groups, with a p-value < 0.0001 and p-value = 0.0006 receptively.

Conclusion

In conclusion, circulating micro-RNA like miR-19b and miR-let7b have a potential opportunity to be a non-invasive diagnostic marker or therapeutic target for PFIC in the future.

ResumenAntecedentes

Los microRNA (miRNA) son un grupo de pequeños RNA no codificantes que se unen al ARNm diana y regulan la expresión génica. Recientemente se han investigado los microRNA circulantes como marcadores de enfermedades y dianas terapéuticas. Aunque varios estudios analizan la expresión de miRNA en enfermedades hepáticas, estos estudios sobre PFIC son escasos. La colestasis intrahepática familiar progresiva (PFIC) es una enfermedad hepática rara con herencia autosómica recesiva. La mayoría de los niños con PFIC progresan a cirrosis e insuficiencia hepática y, en consecuencia, requieren de un trasplante de hígado. El objetivo de este trabajo es la investigación de los niveles de expresión de miR-19b y miR-17b en niños iraníes con PFIC.

Métodos

Se consideraron 25 pacientes con PFIC, 25 niños sanos y 25 pacientes con atresia biliar como grupos de casos y controles. Se obtuvieron muestras de sangre y se midieron las pruebas de función hepática (LFT). Después de la extracción de RNA y la síntesis de cDNA, se realizó PCR cuantitativa usando cebadores específicos para miR-19b y miR-17b. El gen U6 se utiliza como control interno.

Resultados

La qPCR en muestras de pacientes con PFIC demostró que la expresión de miR-19b y miR-17b disminuyó significativamente en los pacientes en comparación con dos grupos de control, con un valor de p < 0,0001 y un valor de p = 0,0006, receptivamente.

Conclusión

En conclusión, los micro-RNA circulantes, como miR-19b y miR-let7b, tienen una oportunidad potencial de ser un marcador de diagnóstico no invasivo o un objetivo terapéutico para PFIC en el futuro.

Section snippetsBackground

Progressive familial intrahepatic cholestasis (PFIC) is a group of heterogeneous pediatric rare disorders with autosomal recessive inheritance caused by a defect in bile secretion and intrahepatic cholestasis that can be leading to death from acute liver failure.1 The prevalence of PFIC is estimated at 1 per 50,000 to 1 per 100,000 births that have been reported from different populations and geographical areas.1 According to the type of mutation, clinical manifestation, liver histology, and

Subjects

This study was performed on blood samples of 25 children who have been diagnosed with PFIC and referred to a cohort clinic affiliated with Shiraz liver transplant research center for enrollment in the Shiraz pediatric liver cirrhosis cohort study (SPLCCS). Diagnosis of PFIC is based on clinical signs and symptoms, routine liver tests, radiographic examinations, and liver biopsy. SPLCCSS was performed on children under 18 years of old with chronic liver disease from all regions of Iran, referred

Results

The median age of the PFIC patients was 1.5 years, range 0.3–12 years. Seventeen (68%) cases were male and the remaining eight (32%) cases were female. The majority of cases (n = 11) were PFIC type 2 (44%) and 8 cases were type1 (32%). The rest of the cases were categorized as type 3 (n = 6, 24%). We ascertained 25 serum samples from healthy children as a control group I and 25 serum samples from BA patients as control group II. The control group 1 were fifteen males (60%) and ten females (40%)

Discussion

Progressive familial intrahepatic cholestasis (PFIC) is an inherited familial cholestatic disease that is caused by defects in biliary secretion.1, 4 The Clinical symptoms usually occur in childhood with progressive cholestasis.1 PFIC is typically progressive and usually leads to cirrhosis and the need for liver transplantation. In the absence of liver transplantation, acute liver failure cause death in childhood. In PFIC-2 hepatocellular carcinoma may develop at a very early age.2, 3

MicroRNAs

Conclusion

In conclusion, our study provides support for the association between miR-19b and miR-let-7 expression in PFIC patients. miR-19b and miR-let-7 levels are significantly reduced in PFIC children in comparison to the control groups (healthy children and BA patients). Circulating levels of miR-19b and miR-let-7 have the potential opportunity to be a biomarker for therapeutic target PFIC in the future but more studies are important to confirm our results.

Authors’ contributions

All of the authors contributed substantially to the concept and design of the study. Material preparation, data collection, and analysis were performed by M.D., N.M., M.H.-A., S.-M.D., B.G., E.E., and N.A. The primary draft of the manuscript was written by M.D. All authors read and approved the final manuscript.

Ethical statement

This study was by the Declaration of Helsinki and approved by the regional ethics committee, IR.SUMS.REC.1398.142.

Consent to participate

Written informed consent was obtained from each patient to take part in the study.

Consent for publication

The authors declare that the personal information of each patient would be confidential. The patients had taken their consent for publication of data before enrolling in the study.

Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Funding

This study was supported by Shiraz University of Medical Science (grant number: 23998).

Conflict of interest

The authors declare no conflict of interest.

Acknowledgment

The authors would like to thank Ms. F. Dara for improving the use of English in the manuscript.

References (27)A. Baker et al.Systematic review of progressive familial intrahepatic cholestasis

Clin Res Hepatol Gastroenterol

(2019)

T.X. Lu et al.MicroRNA

J Allergy Clin Immunol

(2018)

G. Siasos et al.MicroRNAs in cardiovascular disease

Hellenic J Cardiol

(2020)

X. Li et al.miR-19 family: a promising biomarker and therapeutic target in heart, vessels and neurons

Life Sci

(2019)

V. Vinayagamoorthy et al.Newer variants of progressive familial intrahepatic cholestasis

World J Hepatol

(2021)

Morotti RA, Suchy FJ, Magid MS, editors. Progressive familial intrahepatic cholestasis (PFIC) type 1, 2, and 3: a...T. Jones-Hughes et al.Epidemiology and burden of progressive familial intrahepatic cholestasis: a systematic review

Orphanet J Rare Dis

(2021)

S. Vienberg et al.Micro RNA s in metabolism

Acta Physiol

(2017)

X.-M. ChenMicroRNA signatures in liver diseases

World J Gastroenterol

(2009)

J. McDaniel et al.Hyperammonemia results in reduced muscle function independent of muscle mass

Am J Physiol Gastrointest Liver Physiol

(2016)

J. Tsialikas et al.LIN28: roles and regulation in development and beyond

Development

(2015)

Y.-W. Zheng et al.Cellular reprogramming and hepatocellular carcinoma development

World J Gastroenterol

(2013)

J. Balzeau et al.The LIN28/let-7 pathway in cancer

Front Genet

(2017)

View full text

© 2023 Elsevier España, S.L.U. All rights reserved.