Multicenter Observational Study of Chinese Non-Small Cell Lung Cancer (NSCLC) Patients With Rare Driver Gene Mutation

Title: Multicenter observational study for clinicopathological characteristics and clinical efficacy of Chinese Non-Small Cell Lung Cancer (NSCLC) patients With Rare Driver Gene Mutation.

Purpose: To observe the status of rare driver gene mutations in NSCLC patients and identify the subtypes of the mutations.

By comparing and analyzing the relationship between different subtypes, clinicopathological features and clinical efficacy, to find out the effects on anti-tumor therapy and disease survival.

And ultimately to promote the precise application of clinical specifications for new anti-tumor drugs.

Study type: Observational

Inclusion criteria:

1. Female or male, 18 years of age or older；
2. Histologically or cytologically proven diagnosis of NSCLC；
3. Able to get tumor tissue gene testing results by lung cancer Polymerase Chain Reaction(PCR)panel kit carried out in hospital
4. Signed and dated informed consent.

Exclusion criteria:

1. Combine with other tumor type
2. The investigator judges the situation that may affect the clinical search process and results.

Estimated enrollment: 50000 participants.

Outcome measures:

Primary outcome measures：

1. The NSCLC rare driver gene mutation frequency and clinicopathological features in 50,000 patients in real world;
2. The relationship between rare driver gene subtypes and clinicopathological features (age, gender, smoking history, histological subtype, clinical stage, lymph node metastasis, local metastasis, distant metastasis, brain metastasis) in NSCLC patients.

Secondary outcome measures:

1. The relationship between rare driver gene mutation subtypes and disease survival or prognosis (Objective response rate (ORR), Progression-free survival (PFS), and Overall survival (OS)) in NSCLC patients.

Method of Research:

1. Pre- entry/screening period (V0)

   1. Screen the enrolled patients according to the admission criteria. The detection of lung cancer PCR panel kit in the hospital requires the use of tissue samples (including surgical tissue, biopsy tissue under interventional conditions, lymph node biopsy tissue, metastases' tissue, etc.);
   2. All enrolled samples should be tested Neurotrophic-Tropomyosin Receptor Kinase (NTRK) gene mutation (PCR);
   3. If a)+b) results show a rare driver gene mutation was detected, participants will be involved into the next step; if a)+b) test results were negative, then 500 cases were selected for next generation sequencing (NGS) detection in this population, and were involved into the next step;
2. Baseline period (V1)

   a. Do further classification of rare driver gene mutation positive samples by Sanger sequencing, and record the test results;
3. Follow-up period (V2)

   1. Record the disease therapeutic regimen and follow-up of the survival status of the enrolled patients;
   2. Once every 3 months (or according to clinical needs) up to 60 months or death;
   3. Collect information including follow-up treatment, disease status, and survival status, imaging examination results, laboratory examination results, etc. (see the Case Report Form (CRF) form for details);
   4. For those who need further molecular testing (such as primary drug resistance), multi-gene analysis using the next generation sequencing (NGS) method;
   5. Loss of follow-up: If the patient fails to return to the center for a follow-up visit, the center will make two attempts to contact by phone and keep the contact records. If the patient does not respond within 1 month after the second contact, the patient is considered to have lost the interview.

Materials and Methods:

Materials:

The specimen material must be human genomic DNA and total RNA extracted from tumor tissue samples. Before the extraction of DNA and RNA, it is very important to make sure that there is at least 20% tumor cells in the tumor tissue samples.

Methods:

Epidermal Growth Factor Receptor 20 exon insertion (EGFR exon 20-ins) mutation/Activin Receptor-like Kinase (ALK) fusion/ROS proto-oncogene 1 receptor tyrosine kinase (ROS1) fusion/Kirsten Rat Sarcoma Viral Oncogene Homolog (KRAS) mutation/Neuroblastoma RAS viral oncogene homolog (NRAS) mutation/B-Raf proto-oncogene, serine/threonine kinase (BRAF) mutation/Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutation/RET proto-oncogene (RET) fusion/Mesenchymal-Epithelial Transition factor (MET) 14 exon skipping/Erb-b2 Receptor Tyrosine Kinase 2 (ERBB2) mutation/ Neurotrophic-Tropomyosin Receptor Kinase (NTRK) fusion mutation were detected by Fluorogenic Quantitative Polymerase Chain Reaction in Chinese NSCLC.

1. DNA/RNA Extraction:

   The total RNA concentration for gene fusion detection is 10\~100 ng/µL in Formalin-Fixed Paraffin-Embedded (FFPE) tissue or 2\~30 ng/µL in Fresh tissue.

   The amount of extracted DNA for gene mutation detection is 1.5\~3 ng/µL in FFPE tissue or 0.5\~1 ng/µL in Fresh tissue.
2. RNA Reverse Transcription.
3. Detection of the Target Alterations in RNA and DNA:

   ALK, ROS1, RET, NTRK Gene fusion and MET 14 exon skipping mutation were detected in RNA.

   EGFR 20 exon-ins, KRAS, NRAS, BRAF, PIK3CA and ERBB2 mutation were detected in DNA.
4. Result Interpretation

Eligibility Criteria:
Inclusion Criteria:

1. Female or male, 18 years of age or older；
2. Histologically or cytologically proven diagnosis of NSCLC；
3. Able to get tumor tissue gene testing results by lung cancer PCR panel kit carried out in hospital；
4. Signed and dated informed consent。

Exclusion Criteria:

1. Combine with other tumor type
2. The investigator judges the situation that may affect the clinical search process and results