Circulating Tumor Cells and Tumor DNA in HCC and NET

Study Overview
=================
Brief Summary
-----------------
Background Treatment and control of cancer is associated with high costs, to patients in the form of side effects and discomfort during investigations, to society in the form of expensive drugs and studies. Circulating tumor cells (CTC) has received great attention as a cancer biomarker in trying to estimate future course in patients with breast cancer, colon cancer and prostate cancer. CTC is believed to be a crucial step in cancer spreading to the bloodstream and giving rise to metastases. Detection of circulating tumor DNA (ctDNA) specifically adds specificity to the analysis of the CTC. The investigators would like to with molecular biological methods predict which patients requires special monitoring and individualized therapy and explore these tests as clinical decision support. Purpose and method In a blood sample from patients with neuro-endocrine tumor (NET) and hepatocellular carcinoma (HCC), the investigators will by cell separation, flow cytometry and DNA sequencing and digital polymerase chain reaction (PCR): Identify and isolate the CTC and investigate these for tumor-specific mutations. Quantify ctDNA and analyze this for specific mutations, which in the past has been found frequent in NET and HCC. Compare findings of mutations on CTC and ctDNA with mutations in tissue biopsies. The results are compared with the clinical data on disease course, including the effect of treatment and survival. Subjects 40 Patients with small intestinal/unknown primary NET before treatment with somatostatin analogues 30 patients with pancreatic NET before treatment with Everolimus 30 patients with presumed radically treated HCC 30 patients with HCC in treatment with Sorafenib A blood sample will be taken prior to the start of treatment, after 1 month after start of treatment and thereafter every 3.-6. month for up to two years. Perspectives In several cancer types molecular diagnostics have had significant influence in treatment and control strategy. The goal is in future to be able to take advantage of a so-called liquid biopsy as clinical decision support. The study will bring new knowledge to this growing field of research.

Official Title
-----------------
Circulating Tumor Cells and Tumor DNA in HCC and NET - Patient-specific Biomarkers for Clinical Decision Support and Tailored Relapse Diagnostics

Conditions
-----------------
Carcinoma, Hepatocellular, Neuroendocrine Tumors

Intervention / Treatment
-----------------
* Drug: Sorafenib
* Procedure: Radiofrequency ablation (RFA) or surgery
* Drug: Everolimus
* Drug: Lanreotide


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: one of the above mentioned diseases planed surgery, RFA, Somatostatin Analogue, Sorafenib or Everolimus treatment signed informed consent Exclusion Criteria: age below 18, concomitant invasive cancer (not skin cancer) and planed emigration of Denmark.

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 |
| --- | --- |
| HCC sorafenib<br>HCC patients referred for Sorafenib treatment | Drug: Sorafenib<br> <br> * Other names: Anatomical Therapeutic Chemical Classification System L01XE05;|
| HCC curative treatment<br>HCC patient undergoing potential curative treatment, eg. radiofrequency ablation (RFA) or resection | Procedure: Radiofrequency ablation (RFA) or surgery<br>* Intended curative surgery or RFA<br>|
| NET everolimus<br>Pancreatic NET patients referred for Everolimus treatment | Drug: Everolimus<br> <br> * Other names: Anatomical Therapeutic Chemical Classification System L01XE10;|
| NET ssta<br>Small intestinal or unknown primary NET patients referred for treatment with somatostatin analogues, eg. lanreotide and octreotide | Drug: Lanreotide<br>* Or other somatostatin analogues (SSTA), eg. Sandostatin<br>* Other names: Octreotide;|

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

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Concordance between specific DNA mutations found in patient biopsies and plasma circulating tumor DNA | Methods: digital droplet PCR and targeted sequencing of blood samples and biopsies | 2 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Flow cytometry for detection and quantification of CTC in peripheral blood (absolute and relative counts) |  | 3 years | 
| Correlations between mutations found in circulating tumor DNA and amount of circulating tumor cells and treatment response according to RECIST criteria | Methods: digital droplet PCR and targeted sequencing of blood samples, and flowcytometry and cell separation of blood samples | up to 5 years | 
| Correlations between mutations found in circulating tumor DNA and amount of circulating tumor cells and survival | Methods: digital droplet PCR and targeted sequencing of blood samples, and flowcytometry and cell separation of blood samples | up to 5 years | 
| Correlations between mutations fund in circulating DNA and circulating tumor cells | Methods: digital droplet PCR and targeted sequencing of blood samples, and flowcytometry and cell separation of blood samples | 3 years | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
biopsy, tumor DNA, circulating tumor cells