The Red Blood Cells Based Blood Test for Lung Cancer EARLY Detection

Study Overview
=================
Brief Summary
-----------------
This study plan to enroll 852 patients with pulmonary nodules smaller than 3 cm in diameter, whose DNA and RNA will be extracted from mature red blood cells isolated from peripheral blood. The DNA 5-methylcytosine(5-mC) and RNA 2'-O-methylation information will be acquired by NGS and Nm Judge Universally sequencing (NJU-Seq), which will be used to establish models to distinguish patients with benign and malignant nodule in the training group and further evaluated in the validation group. The pathological results will be acquired after surgery or biopsy as standard in the study.

Detailed Description
-----------------
This is a prospective, multi-center, observational, cohort study and seeks to enroll 852 participants with pulmonary nodules smaller than 3 cm in diameter from 4 hospitals in China. Each participant pulmonary nodule is ≤30mm as assessed by CT scanning, nodule is suspected to be malignant, and plan to use biopsy or surgery for diagnosis or treatment within 60 days.Their blood samples, CT scan data, and clinical data will be collected at each visit and tissues will be collected when participants who receive pneumonectomy or percutaneous lung biopsy. Each subject will analysis DNA and RNA methylation in nucleic acids extracted from mature red blood cells isolated from peripheral blood by next-generation sequencing (NGS) . A proprietary algorithm will be used to identify the modification sites and pattern for differentiating the benign and malignant pulmonary nodules. Sensitivity and specificity of the different methylation panels will be to evaluated.

Official Title
-----------------
The Training and Validation Study: Development of a Red Blood Cells Based Blood Test for Early Detection of Lung Cancer

Conditions
-----------------
Lung Cancer, Red Blood Cells, Methylation

Intervention / Treatment
-----------------
* Diagnostic Test: Methylation tests of nucleic acids extracted from mature red blood cells (RBCs)


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Pulmonary nodule is ≤3cm; Subjects suspected of lung cancer; Diagnosis or treatment by biopsy or surgery within 60 days; Signed informed consent. Exclusion Criteria: History of malignant tumors; Received anti-tumor therapy; Can't collect the histopathological results or the pathological results are metastatic cancer; Can't obtain detection information of red blood cells; Pregnancy or lactating female； Other situations not suitable for the study.

Ages Eligible for Study
-----------------
Minimum Age: 18 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
No

Study Plan
=================
How is the study designed?
-----------------

Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Patients with pulmonary nodules (diameter ≤3cm) plan to diagnosis benign and malignant<br>Patients with pulmonary nodules (diameter ≤3cm) plan to use biopsy or surgery or treatment to diagnosis benign and malignant. | Diagnostic Test: Methylation tests of nucleic acids extracted from mature red blood cells (RBCs)<br>* Up to 15 ml of peripheral blood will be collected from each subject, and the blood specimen will be processed to isolate mature red blood cells and extract the DNA and RNA from RBCs. DNA and RNA methylation will be tested by next-generation sequencing (NGS).<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sensitivity of the RBCs-based blood test for differentiating benign and malignant pulmonary nodules diameter less than 3 cm using DNA methylation analysis by NGS. |  | 3 years | 
| Specificity of the RBCs-based blood test for differentiating benign and malignant pulmonary nodules diameter less than 3 cm using DNA methylation analysis by NGS. |  | 3 years | 
| Positive predictive value (PPV) of the RBCs-based blood test for differentiating benign and malignant pulmonary nodules diameter less than 3 cm using DNA methylation analysis by NGS. |  | 3 years | 
| Negative predictive value (NPV) of the RBCs-based blood test for differentiating benign and malignant pulmonary nodules diameter less than 3 cm using DNA methylation analysis by NGS. |  | 3 years | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sensitivity of the RBCs-based methylation assay in the invasion of malignant pulmonary nodules. |  | 3 years | 
| Specificity of the RBCs-based methylation assay in the invasion of malignant pulmonary nodules. |  | 3 years | 
| Area under curve (AUC) of receiver operating characteristic curve (ROC) in the model established in this study will compare with that of Mayo Clinic and Veterans Affairs models. |  | 3 years | 
| Sensitivity in the model established in this study will compare with that of Mayo Clinic and Veterans Affairs models. |  | 3 years | 
| Specificity in the model established in this study will compare with that of Mayo Clinic and Veterans Affairs models. |  | 3 years |