Single-cell Sequencing and Establishment of Models in Neuroendocrine Neoplasm

Study Overview
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Brief Summary
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The aim of this study is to use single-cell sequencing technology to explore neuroendocrine neoplasm (NEN) molecular biological characteristics, tumor heterogeneity and cell subtypes. Besides. NEN models are constructed for basic research, including primary cell lines, organoids, and animal models.

Detailed Description
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Neuroendocrine neoplasm (NEN) generally refers to all tumors that originate from peptidergic neurons and neuroendocrine cells. It is a relatively rare tumor, ranging from indolent, slow-growing low-grade malignant tumors to obvious distant metastases. A series of malignant tumors. The most common place for neuroendocrine tumors is in the digestive system, and about two-thirds of neuroendocrine tumors occur in the gastrointestinal pancreas. However, with the change of people's living habits and the improvement of physical examination awareness, the incidence of neuroendocrine tumors increased from 1.09/100,000 in 1973 to 6.98/100,000 in 2012. The incidence has increased by 6 times compared with other tumors. , NENs increase more rapidly. The incidence of people over 65 years of age has increased by 8 times, and the incidence of people younger than 50 years of age has also increased by 3 times, and the incidence has been increasing year by year. It is worth noting that the average age of pancreatic neuroendocrine tumors is only 56.7-13.3 years old, showing a younger trend. Because most neuroendocrine tumors lack specific manifestations, coupled with unique inert biological characteristics, clinicians often lack understanding, which can easily lead to misdiagnosis and treatment. Taking gastrointestinal pancreatic neuroendocrine tumors (GEP-NEN) as an example, due to the uneven diagnosis and treatment level, GEP-NEN patients may be diagnosed with a delay of up to 7 years. Because of this, as many as 40-95% of GEP-NEN patients have developed distant metastases at the time of diagnosis, and 65-95% of them have liver metastases. Liver metastasis is the most critical prognostic risk factor in GEP-NEN, and it has a decisive impact on the survival of patients. The 5-year survival rate of gastrointestinal neuroendocrine tumors with liver metastasis is 56%-83%, while the 5-year survival rate of pancreatic neuroendocrine tumors is only 48.8%, and the 10-year survival rate is only 30.2%. For limited-stage patients, even if radical surgical resection, up to 94% of cases will still have recurrence and metastasis within 5 years. For inoperable patients, although chemotherapy, targeted therapy, biological therapy and other methods can be used, the effect is still limited. Therefore, the systematic development of basic-clinical translational research on neuroendocrine tumors is very important and imminent. Based on the above-mentioned problems, this project intends to conduct a systematic and in-depth study of neuroendocrine tumors: 1. Use single-cell sequencing technology to deeply study the molecular biological characteristics of NEN tumors, tumor heterogeneity and cell subtypes. 2. Construct NEN models, including primary cell lines, organoids, and mouse animal models. This study can establish a neuroendocrine tumor research system to find the molecular mechanism and potential intervention targets of NEN recurrence and metastasis, and provide clinicians with safe and effective treatment strategies, thereby improving the therapeutic effect of neuroendocrine tumors.

Official Title
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Single-cell Sequencing and Establishment of Cell Lines, Organoids, and Transplanted Tumor Models in Neuroendocrine Neoplasm

Conditions
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Neuroendocrine Neoplasm

Intervention / Treatment
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* Procedure: Biopsy, open or laparoscopic surgery


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Between the ages of 18 and 80, with any gender; Physical fitness score ECOG 0～1 points; The diagnosis is considered as gastrointestinal pancreatic neuroendocrine tumor or pancreatic ductal adenocarcinoma; There is no obvious contraindication to surgery or biopsy; Uncompensated liver cirrhosis, acute and chronic hepatitis and other diseases; No history of other biliary tract related diseases; Volunteer to participate and sign the informed consent form. - Exclusion Criteria: Patients with non-gastrointestinal pancreatic neuroendocrine tumors or pancreatic ductal adenocarcinoma confirmed pathologically; Suffer from other digestive system diseases, such as irritable bowel syndrome, inflammatory bowel disease; Those who have incomplete follow-up data or refuse to accept follow-up; Patients with other malignant tumors or hematological diseases; -

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 80 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Gastroenteropancreatic neuroendocrine neoplasms<br>Collect biopsy/surgical fresh tissue of gastroenteropancreatic neuroendocrine neoplasms. | Procedure: Biopsy, open or laparoscopic surgery<br>* Collect NEN and PDAC biopsy or surgical fresh tissue to conduct single cell sequencing or model construction.<br>|
| Pancreatic ductal adenocarcinoma<br>Collect biopsy/surgical fresh tissue of pancreatic ductal adenocarcinoma. | Procedure: Biopsy, open or laparoscopic surgery<br>* Collect NEN and PDAC biopsy or surgical fresh tissue to conduct single cell sequencing or model construction.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Bioinformatics analysis of single-cell sequencing results | Analysis of NEN Molecular Biology Information Using Single Cell Sequencing Technology | Half a year | 
| Model construction | Collect fresh specimens of NEN to cultivate primary cell lines, construct organoids, and establish animal models of NEN | One year | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Exploration of NEN organoid | To explore and optimize the medium scheme of NEN orgnoid and methods of histomorphological identification of type organs | One year | 
| Tumor microenvironment | The tumor microenvironment of NEN was analyzed | One year | 
| Comparison of mechanism of NEN with PDAC | Collect fresh specimens of pancreatic ductal adenocarcinoma (PDAC) for single-cell sequencing and organoid construction for comparison with NEN | Half a year | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Patient-derived Tumor Xenograft, Neuroendocrine Neoplasm, Single Cell Sequencing Technology, Primary Cell Culture, Organoid