Identification and Characterization of Circulating microRNAs as Diagnostic Biomarkers in Adolescent Idiopathic Scoliosis
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 入组人数
- 30
- 试验地点
- 2
- 主要终点
- Isolation of circulating microRNAs from AIS patients and healthy controls
研究概览
简要总结
The aim of the present study is to evaluate the expression of a large panel of microRNAs, already known and validated in other ortopedic pathologies and bone metabolism, in the plasma of Adolescent Idiopathic Scoliosis (AIS) patients. The deregulated microRNAs identified will be then validated and computational analyzes will determine their potential involvement in the metabolism of bone and/or cartilage tissue in order to correlate the results obtained with the clinical data of the AIS patients. The investigators aimed to develop a microRNAs panel to further validate in a larger population of AIS patients in order to produce a device for the diagnosis and prognosis of Molecular-based AIS.
详细描述
MATERIALS AND METHODS: 1) Collection of a venous blood sample from patients enrolled with AIS or controls, and subsequent separation of plasma; 2) Isolation of circulating total RNA from the plasma of AIS patients (Kit Invitrogen: Total RNA Isolation Reagent) and preparation of microRNA samples (TaqMan™ MicroRNA Reverse Transcription Kit); 3) Analysis of microRNA expression profiles by micro-array technique (Taqman Advanced miRNAs Human CardA + B); 4) Validation of deregulated microRNAs by single-qRT-PCR assay and computational functional analyses; and 5) In vitro studies to investigate the molecular role of the microRNAs identified within the mechanisms of bone remodeling.
Enrollment Procedures:
Patients considered eligible will be enrolled in the study, after providing a written informed consent. Healthy subjects (control group) will be recruited, on a voluntary basis, by means of an announcement published on the web page of the Rizzoli Orthopedic Institute (www.ior.it), "News" section.
Data Collection: Clinical data will be retrieved by patient's source document. A protocol-specific CRF reporting the results of the analyses will be provided. A CRF is required and should be completed for each included subject.
Ethics: The clinical trial protocol and its documents will be sent before initiating the study to the competent Authorities and Ethics Committees of each participating country for its approval. The responsible investigator will ensure that this study is conducted in agreement with either the most updated Declaration of Helsinki and all the international and local laws that apply to clinical trials and to patient protection. The protocol has been written, and the study will be conducted according to the principles of the ICH Harmonized Tripartite Guideline for Good Clinical Practice (ref: http://www.emea.eu.int/pdfs/human/ich/013595en.pdf).
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Other
入排标准
- 年龄范围
- 11 Years 至 17 Years（Child）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Age between 11-17 years
- •Diagnosis of idiopathic scoliosis with a Cobb angle> 10 °
- •Minimum follow-up of two years
- •Clinical data and radiological tests available and no surgical treatment prior to enrollment in the study
- •Age between 11-17 years
- •Healthy subjects not affected by orthopedic and oncological diseases
排除标准
- •Severe cognitive impairment or psychiatric disorders
- •Patients with scoliosis due to secondary causes
- •Women of childbearing age must not be pregnant
结局指标
主要结局
Isolation of circulating microRNAs from AIS patients and healthy controls
时间窗: Within 24 hours of blood sampling
Plasma samples will be obtained from blood samples of 20 AIS patients and 10 healthy subjects, collected in commercially available anticoagulant-treated tubes and centrifuged for 15 minutes at 2,000 x g. Circulating microRNAs will be isolated from the plasma samples by using commercially available miRNA Plasma Purification kit, specifically designed for purify high-quality microRNAs from plasma samples. Quantification and quality control of the purified circulating microRNAs will be determined through micro-spectrophotometry and microfluidics-based platforms.
Expression profiling of circulating microRNAs from AIS patients and healthy controls
时间窗: Within 2 months of blood sampling
The differentially expressed circulating microRNAs between AIS patients and healthy controls will be screened through micro-array technology using commercially available custom array microRNA cards, developed for profiling hundreds of unique human mature microRNAs known to be present in serum/plasma samples.
次要结局
- Bioinformatics analysis of circulating microRNAs in AIS patients and healthy controls.(Within 3 months of blood sampling)
- Functional role of the circulating microRNAs related to AIS(Within 6 months of blood sampling)