The structural integrity of mitochondrial DNA reveals new molecular perspectives for early diagnosis of colorectal cancer

The structural integrity of mitochondrial DNA reveals new molecular perspectives for early diagnosis of colorectal cancer - mtDNA-KRK

Status

Active, not recruiting

Phases

Unknown

Study type

Observational

Source

DRKS

Registry ID

DRKS00030257

Enrollment

Registered

2022-09-29

Start date

2021-02-02

Completion date

Unknown

Last updated

2025-04-07

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

C18 C20

Interventions

Group 1: The project aims to: Statistically compare the fragment length of cell-free DNA in the blood plasma of 80 patients with a confirmed diagnosis of colorectal carcinoma in stages I-IV with the

Eligibility

Sex/Gender

All

Age

18 Years to No maximum

Inclusion criteria

Inclusion criteria: 1. Healthy subjects (must be available for purchase for research purposes (at Central Biobank GmbH) between the ages of 18 and 90 2. All male and female colorectal cancer patients (> 18 years) are included with the ability to understand and question the relevant patient information with a diagnosis of colorectal cancer who are referred for surgery 3. Patient's willingness to provide blood and tissue samples for scientific analysis as part of the preliminary study 4. A written declaration of consent (for healthy people and patients) must be available

Exclusion criteria

Exclusion criteria: 1. Healthy subjects under 18 years of age 2. Colorectal cancer patients with manifest coronary heart disease, chronic heart failure and peripheral arterial disease 3. Colorectal cancer patients with other tumor diseases 4. Colon cancer patients with advanced renal failure 5. Colon cancer patients with chronic inflammatory diseases 6. Healthy subjects and patients who do not meet the inclusion criteria or who their Withdraw consent to participate in the study

Design outcomes

Primary

Measure	Time frame
The aim of this study is: 1) to analyze total cfDNA concentration in CRC patients with different histopathological stages (UICC I-IV). 2) to address the question whether the integrity index of cfDNA increases in CRC patients as a result of elevated necrotic degradation processes. Therefore, we intended to quantify short and long fragments of cfDNA via qPCR independent from total cfDNA levels by using equal template concentrations for CRC patients and healthy individuals. This approach may help to understand whether previously described diagnostic markers are either increased, decreased or unaltered in blood plasma of CRC patients compared to healthy individuals.	—

Measure

Time frame

The aim of this study is: 1) to analyze total cfDNA concentration in CRC patients with different histopathological stages (UICC I-IV). 2) to address the question whether the integrity index of cfDNA increases in CRC patients as a result of elevated necrotic degradation processes. Therefore, we intended to quantify short and long fragments of cfDNA via qPCR independent from total cfDNA levels by using equal template concentrations for CRC patients and healthy individuals. This approach may help to understand whether previously described diagnostic markers are either increased, decreased or unaltered in blood plasma of CRC patients compared to healthy individuals.

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Secondary

Measure	Time frame
Patients and samples: Blood samples of 80 consecutive patients were collected before surgical care. The CRC cohort included patients with UICC stage I (n=21), UICC II (n=21), UICC III (n=20) and UICC IV (n=18), confirmed after surgery by histopathological examination according to established standard diagnostic procedures (gold standard). Blood samples from 50 healthy individuals were provided by Central BioHub®, a commercial Biobank that hosts collections of human biospecimen for scientific research, served as controls. All blood samples were collected using K2EDTA BD Vacutainer® Collection Tube (Becton Dickinson, Germany). On the day of venipuncture, plasma samples were centrifuged at 2,000 x g for 10 min and the supernatants were carefully removed, avoiding the buffy-coat. Plasma aliquots of CRC patients and controls were stored at –80°C until analysis. Sample processing, total cfDNA extraction and quantification: A total of 1 ml of plasma samples from all individuals were thawed at room temperature and centrifuged at 16,000 x g for 10 minutes at 4°C. The supernatant was transferred to 1.5 ml tube and cfDNA extraction performed using the QIAamp® Circulating Nucleic Acid Kit (Qiagen, Germany) according to the manufacturer´s protocol, except the column-based isolation of total cfDNA, that was performed with centrifugation at 2,000 g instead of using the vacuum pump. Extracted cfDNA was eluted with 50 µl AVE buffer and total cfDNA concentration was determined using Qubit™ dsDNA HS Assay Kit and Qubit™ 3.0 Fluorometer (ThermoFisher, Fisher Scientific, Invitrogen, Germany). The samples were stored at –20°C prior to quantitative real-time PCR (qPCR) analysis. Quantity and integrity index of nuclear and mitochondrial cfDNA: To measure n-cfDNA and mt-cfDNA concentrations presumably originating either from apoptotic or necrotic cell death, short and long fragments of KRAS, Alu and MTCO3 markers were targeted with qPCR. Note, that n-cfDNA markers in the plas	—

Measure

Time frame

Patients and samples: Blood samples of 80 consecutive patients were collected before surgical care. The CRC cohort included patients with UICC stage I (n=21), UICC II (n=21), UICC III (n=20) and UICC IV (n=18), confirmed after surgery by histopathological examination according to established standard diagnostic procedures (gold standard). Blood samples from 50 healthy individuals were provided by Central BioHub®, a commercial Biobank that hosts collections of human biospecimen for scientific research, served as controls. All blood samples were collected using K2EDTA BD Vacutainer® Collection Tube (Becton Dickinson, Germany). On the day of venipuncture, plasma samples were centrifuged at 2,000 x g for 10 min and the supernatants were carefully removed, avoiding the buffy-coat. Plasma aliquots of CRC patients and controls were stored at –80°C until analysis. Sample processing, total cfDNA extraction and quantification: A total of 1 ml of plasma samples from all individuals were thawed at room temperature and centrifuged at 16,000 x g for 10 minutes at 4°C. The supernatant was transferred to 1.5 ml tube and cfDNA extraction performed using the QIAamp® Circulating Nucleic Acid Kit (Qiagen, Germany) according to the manufacturer´s protocol, except the column-based isolation of total cfDNA, that was performed with centrifugation at 2,000 g instead of using the vacuum pump. Extracted cfDNA was eluted with 50 µl AVE buffer and total cfDNA concentration was determined using Qubit™ dsDNA HS Assay Kit and Qubit™ 3.0 Fluorometer (ThermoFisher, Fisher Scientific, Invitrogen, Germany). The samples were stored at –20°C prior to quantitative real-time PCR (qPCR) analysis. Quantity and integrity index of nuclear and mitochondrial cfDNA: To measure n-cfDNA and mt-cfDNA concentrations presumably originating either from apoptotic or necrotic cell death, short and long fragments of KRAS, Alu and MTCO3 markers were targeted with qPCR. Note, that n-cfDNA markers in the plas

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Countries

Germany

Contacts

Public ContactRené Mantke

Klinik für Allgemein und Viszeralchirurgie

chirurgie@klinikum-brandenburg.de03381411200

Outcome results

None listed

Source: DRKS (via WHO ICTRP) · Data processed: Feb 11, 2026