DiabetesCEC4DCNCT05169502
Discovering Early Biomarkers in Circulating Endothelial Cells for Diabetes Complications by Single Cell RNA Sequencing
An observational study of Diabetes and Diabetes Mellitus, run by Aarhus University Hospital at 1 site in Denmark.
Trial summary
The purpose of this study is to discover early biomarkers in circulating endothelial cells for diabetes complications, by investigating circulating endothelial cells in blood samples from patients with newly diagnosed proliferative diabetic retinopathy, newly diagnosed maculopathy, patients with diabetes without eye… Read more about this study
- Who is it for?
- Ages 18 years and olderHealthy volunteers can take part
- Conditions the study is for
- Diabetes, Diabetes Mellitus, Proliferative Diabetic Retinopathy
- How many people
- 40 participants
- Where
- 1 siteDenmark
- Study dates
- 8 Dec 2021to 12 Aug 2024
- How long will I take part?
- 1 Day
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 years and older
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
Study population: The patients with newly diagnosed proliferative diabetic retinopathy and patients with newly diagnosed diabetic maculopathy will be recruited at department of Ophthalmology at Aarhus University Hospital. The healthy individuals and patients with diabetes and without eye diseases will be recruited at Steno Diabetes Center Aarhus, Aarhus University Hospital.
Who may be able to join
- Type I or II diabetes
- Habile
- Age> 18 years
Who cannot join
- Incapacitated
- Age < 18 years
Trial locations and contacts
1 site in 1 country.
Denmark1 site
Steno Diabetes Center Aarhus, Aarhus University Hospiral
Aarhus, 8000
About this study
Official title: Identification of Biomarkers for Early Detection of Diabetes Complications by Characterisation of Circulating Endothelial Cells in Human Blood Using Single-cell RNA Sequencing.
The purpose of this study is to discover early biomarkers in circulating endothelial cells for diabetes complications, by investigating circulating endothelial cells in blood samples from patients with newly diagnosed proliferative diabetic retinopathy, newly diagnosed maculopathy, patients with diabetes without eye diseases, and individuals without diabetes by single-cell RNA sequencing. The single-cell RNA sequencing analysis will make it possible to fully phenotype diabetes circulating endothelial cells at single-cell level and reveal the first atlas of circulating endothelial cells in humans at both healthy and diabetes conditions.
Read the detailed description
There is one particular type of ECs, which are circulating in the blood system called circulating endothelial cells (CECs). The common concept of CECs only refers to mature ECs that have been shed from the vascular wall due to impaired vascular functions caused by e.g. diabetes, and hypertension. Mature CECs have thus been proposed as highly valuable targets for diagnosis, treatment, and prognosis of cardiovascular diseases. In healthy individuals, the number of CECs varies from 0 to 7000 per ml of blood and increases significantly in patients with hypertension, diabetes, and cardiovascular diseases. In addition to mature EC, which circulates as CEC. Immature EC, called "circulating endothelial progenitor cells" (cEPC), is reduced in patients with cardiovascular disorders. Based on the highly heterogeneous phenotype of vascular ECs found within and between tissues, it is not surprising that all currently known surface markers for both mature CEC and cEPC are debatable as these studies did not consider ECs heterogeneity. Proliferative diabetic retinopathy (PDR) is a well-defined diabetes complication caused by ECs dysfunction. This study will examine human blood (from healthy individuals, patients with diabetes and PDR or diabetic maculopathy, or without eye disease) and thereby characterize both CEC and cEPC by single-cell RNA sequencing.
This study aims to discover early CEC biomarkers for diabetes complications exemplified with diabetic eye diseases.
Recruitment and scRNA-seq: Blood samples from healthy individuals and patients with diabetes and PDR, diabetic maculopathy, or without eye disease will be recruited from Steno Diabetes Center Aarhus and the Department of Ophthalmology at Aarhus University Hospital. The PBMCs will be freshly isolated from the blood samples followed by CEC enrichment by Fluorescent-Activated Cell Sorting of CD45-CD31+ live cells. Ethical and GDPR approval has been obtained.
ScRNA-seq of CECs: The CD45-CD31+ cells will be processed immediately to single-cell capture and -barcoded cDNA synthesis using the Next GEM Single Cell 3' GEM kit (10x genomics). NGS libraries will be prepared and sequenced using the MGI2000 sequencers from BGI. GenomeDK will be used to supercomputing clusters followed by scRNA-seq analysis and visualization. Integrative multi-model scRNA-seq analyses: Standard scRNA-seq analyses will be carried out to stratify the differential subtypes of CECs and cEPCs and identify ECs subtypes associated with the development of proliferative diabetic retinopathy and diabetic maculopathy. Integrative analysis of scRNA-seq results aims to discover CECs biomarkers which can be used for early diagnosis, prevention and treatment of diabetes complications.
- Conditions
- Diabetes, Diabetes Mellitus, Proliferative Diabetic Retinopathy, Diabetic Maculopathy, Circulating Endothelial Cells
- Medical subject headings
- Diabetes Mellitus
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
| Study group | Role | What they receive |
|---|---|---|
| Patients with diabetes and newly diagnosed proliferative diabetic retinopathy, level 4. (n=10) | Inclusion criteria: Type I or II diabetes, habile and age>18 years. Exclusion criteria: Incapacitated or age< 18 years. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing. A questionnaire describing the patients general health (gender, age, duration of diabetes, smoking status, blood pressure, BMI and hip/waist ratio) will be made. | |
| Patients with diabetes and newly diagnosed diabetic maculopathy. (n=10) | These patients will be matched 1:1 to the patients with proliferative diabetic retinopathy, regarding gender, age, duration of diabetes, smoking status, and blood pressure. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing.A questionnaire describing the patients general health (gender, age, duration of diabetes, smoking status, blood pressure, BMI and hip/waist ratio) will be made. | |
| Patients with diabetes without retinopathy, level 0. (n=10) | These patients will be matched 1:1 to the patients with proliferative diabetic retinopathy, regarding gender, age, duration of diabetes, smoking status, and blood pressure. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing.A questionnaire describing the patients general health (gender, age, duration of diabetes, smoking status, blood pressure, BMI and hip/waist ratio) will be made. | |
| Individuals without diabetes and known eye diseases (n=10) | These patients will be matched 1:1 to the patients with proliferative diabetic retinopathy, regarding gender, age, smoking status, and blood pressure. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing.A questionnaire describing the patients general health (gender, age, smoking status, blood pressure, BMI and hip/waist ratio) will be made. |
What is being measured
The primary outcome is the main question the study is designed to answer.
Genome wide expression profile of circulating cells in blood samples from each cohort group
Flow cytometry and single-cell RNA sequencing will be used to identify and consolidate potential biomarkers for diabetes complications, by making a genome wide expression profile used to characterise the different phenotypes of circulating cells in each cohort group.
Measured: The PBMCs (including circulating endothelial cells) will be freshly isolated from the whole blood sampel, and frozen until all samples are collected (8 months). Afterwards flow cytometry and scRNA-seq analysis will be performed on all samples (2 months).
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted27 Dec 2021
- Study start8 Dec 2021
- Primary completion1 Aug 2024
- Study completion12 Aug 2024
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (4)Papers the sponsor has linked to this study.
- Other trials for DiabetesStudies of the same condition, open and under way.
- More trials run by Aarhus University HospitalThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Prospective
- Enrolment
- 40 participants (actual)
- Target duration
- 1 Day
Sponsor and investigators
- Lead sponsor
- Aarhus University Hospital
- Sponsor type
- University, hospital or other
- Collaborators
- University of Aarhus
- Responsible party
- Principal investigator, Camilla Blunk Brandt, Aarhus University Hospital
- Investigators
- Niels Jessen, Professor, Principal investigator, Steno Diabetes Center Aarhus, Aarhus Universitet Hospital
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT05169502
- Sponsor's protocol ID
- 33579
- Other IDs
- AUFF-NOVA Research Grant
- Acronym
- CEC4DC
- Status last verified
- Aug 2024
- First submitted
- 24 Nov 2021
- Last updated
- 20 Aug 2024
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Yes: This project will reveal the first atlas (open acces) of circulating endothelial cells in human at both healthy and diabetes conditions.
Publications
Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G, Nishigaki I. The vascular endothelium and human diseases. Int J Biol Sci. 2013 Nov 9;9(10):1057-69. doi: 10.7150/ijbs.7502. eCollection 2013.
PubMed 24250251Carmeliet P. Angiogenesis in health and disease. Nat Med. 2003 Jun;9(6):653-60. doi: 10.1038/nm0603-653.
PubMed 12778163Damani S, Bacconi A, Libiger O, Chourasia AH, Serry R, Gollapudi R, Goldberg R, Rapeport K, Haaser S, Topol S, Knowlton S, Bethel K, Kuhn P, Wood M, Carragher B, Schork NJ, Jiang J, Rao C, Connelly M, Fowler VM, Topol EJ. Characterization of circulating endothelial cells in acute myocardial infarction. Sci Transl Med. 2012 Mar 21;4(126):126ra33. doi: 10.1126/scitranslmed.3003451.
PubMed 22440735Aird WC. Endothelial cell heterogeneity. Cold Spring Harb Perspect Med. 2012 Jan;2(1):a006429. doi: 10.1101/cshperspect.a006429.
PubMed 22315715
Study at a glance
From the registry record, last updated by the sponsor on 20 Aug 2024. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Identification of Biomarkers for Early Detection of Diabetes Complications by Characterisation of Circulating Endothelial Cells in Human Blood Using Single-cell RNA Sequencing.
- ClinicalTrials.gov ID
- NCT05169502
- Acronym / sponsor ID
- CEC4DC / 33579
- Conditions
- Diabetes, Diabetes Mellitus, Proliferative Diabetic Retinopathy, Diabetic Maculopathy, Circulating Endothelial Cells
- Study type
- Observational
- Recruitment status
- Completed
- Enrolment
- 40 participants (actual)
- Lead sponsor
- Aarhus University HospitalUniversity, hospital or other
- Collaborators
- University of Aarhus
- Study start
- 8 Dec 2021 (actual)
- Primary completion
- 1 Aug 2024 (actual)
- Study completion
- 12 Aug 2024 (actual)
- First posted
- 27 Dec 2021 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
The purpose of this study is to discover early biomarkers in circulating endothelial cells for diabetes complications, by investigating circulating endothelial cells in blood samples from patients with newly diagnosed proliferative diabetic retinopathy, newly diagnosed maculopathy, patients with diabetes without eye diseases, and individuals without diabetes by single-cell RNA sequencing. The single-cell RNA sequencing analysis will make it possible to fully phenotype diabetes circulating endothelial cells at single-cell level and reveal the first atlas of circulating endothelial cells in humans at both healthy and diabetes conditions.
Detailed description
There is one particular type of ECs, which are circulating in the blood system called circulating endothelial cells (CECs). The common concept of CECs only refers to mature ECs that have been shed from the vascular wall due to impaired vascular functions caused by e.g. diabetes, and hypertension. Mature CECs have thus been proposed as highly valuable targets for diagnosis, treatment, and prognosis of cardiovascular diseases. In healthy individuals, the number of CECs varies from 0 to 7000 per ml of blood and increases significantly in patients with hypertension, diabetes, and cardiovascular diseases. In addition to mature EC, which circulates as CEC. Immature EC, called "circulating endothelial progenitor cells" (cEPC), is reduced in patients with cardiovascular disorders. Based on the highly heterogeneous phenotype of vascular ECs found within and between tissues, it is not surprising that all currently known surface markers for both mature CEC and cEPC are debatable as these studies did not consider ECs heterogeneity. Proliferative diabetic retinopathy (PDR) is a well-defined diabetes complication caused by ECs dysfunction. This study will examine human blood (from healthy individuals, patients with diabetes and PDR or diabetic maculopathy, or without eye disease) and thereby characterize both CEC and cEPC by single-cell RNA sequencing.
This study aims to discover early CEC biomarkers for diabetes complications exemplified with diabetic eye diseases.
Recruitment and scRNA-seq: Blood samples from healthy individuals and patients with diabetes and PDR, diabetic maculopathy, or without eye disease will be recruited from Steno Diabetes Center Aarhus and the Department of Ophthalmology at Aarhus University Hospital. The PBMCs will be freshly isolated from the blood samples followed by CEC enrichment by Fluorescent-Activated Cell Sorting of CD45-CD31+ live cells. Ethical and GDPR approval has been obtained.
ScRNA-seq of CECs: The CD45-CD31+ cells will be processed immediately to single-cell capture and -barcoded cDNA synthesis using the Next GEM Single Cell 3' GEM kit (10x genomics). NGS libraries will be prepared and sequenced using the MGI2000 sequencers from BGI. GenomeDK will be used to supercomputing clusters followed by scRNA-seq analysis and visualization. Integrative multi-model scRNA-seq analyses: Standard scRNA-seq analyses will be carried out to stratify the differential subtypes of CECs and cEPCs and identify ECs subtypes associated with the development of proliferative diabetic retinopathy and diabetic maculopathy. Integrative analysis of scRNA-seq results aims to discover CECs biomarkers which can be used for early diagnosis, prevention and treatment of diabetes complications.
Study design
- Observational model
- Cohort
- Time perspective
- Prospective
- Target follow-up
- 1 Day
- Number of arms
- 4
| Arm | Type | Receives |
|---|---|---|
| Patients with diabetes and newly diagnosed proliferative diabetic retinopathy, level 4. (n=10) | – | –Inclusion criteria: Type I or II diabetes, habile and age>18 years. Exclusion criteria: Incapacitated or age< 18 years. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing. A questionnaire describing the patients general health (gender, age, duration of diabetes, smoking status, blood pressure, BMI and hip/waist ratio) will be made. |
| Patients with diabetes and newly diagnosed diabetic maculopathy. (n=10) | – | –These patients will be matched 1:1 to the patients with proliferative diabetic retinopathy, regarding gender, age, duration of diabetes, smoking status, and blood pressure. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing.A questionnaire describing the patients general health (gender, age, duration of diabetes, smoking status, blood pressure, BMI and hip/waist ratio) will be made. |
| Patients with diabetes without retinopathy, level 0. (n=10) | – | –These patients will be matched 1:1 to the patients with proliferative diabetic retinopathy, regarding gender, age, duration of diabetes, smoking status, and blood pressure. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing.A questionnaire describing the patients general health (gender, age, duration of diabetes, smoking status, blood pressure, BMI and hip/waist ratio) will be made. |
| Individuals without diabetes and known eye diseases (n=10) | – | –These patients will be matched 1:1 to the patients with proliferative diabetic retinopathy, regarding gender, age, smoking status, and blood pressure. One 20mL blood sample from each patient will be analysed by flow cytometry and single cell RNA sequencing.A questionnaire describing the patients general health (gender, age, smoking status, blood pressure, BMI and hip/waist ratio) will be made. |
Who may qualify
- Ages
- 18 years and older
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Adult, Older adult
- Study population
- The patients with newly diagnosed proliferative diabetic retinopathy and patients with newly diagnosed diabetic maculopathy will be recruited at department of Ophthalmology at Aarhus University Hospital. The healthy individuals and patients with diabetes and without eye diseases will be recruited at Steno Diabetes Center Aarhus, Aarhus University Hospital.
Key inclusion criteria (3 listed)
- Type I or II diabetes
- Habile
- Age> 18 years
Key exclusion criteria (2 listed)
- Incapacitated
- Age < 18 years
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Primary outcome
Genome wide expression profile of circulating cells in blood samples from each cohort group
Time frame: The PBMCs (including circulating endothelial cells) will be freshly isolated from the whole blood sampel, and frozen until all samples are collected (8 months). Afterwards flow cytometry and scRNA-seq analysis will be performed on all samples (2 months).
Flow cytometry and single-cell RNA sequencing will be used to identify and consolidate potential biomarkers for diabetes complications, by making a genome wide expression profile used to characterise the different phenotypes of circulating cells in each cohort group.
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 20 Aug 2024.
- Background: PubMed 24250251Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G, Nishigaki I. The vascular endothelium and human diseases. Int J Biol Sci. 2013 Nov 9;9(10):1057-69. doi: 10.7150/ijbs.7502. eCollection 2013.
- Background: PubMed 12778163Carmeliet P. Angiogenesis in health and disease. Nat Med. 2003 Jun;9(6):653-60. doi: 10.1038/nm0603-653.
- Background: PubMed 22440735Damani S, Bacconi A, Libiger O, Chourasia AH, Serry R, Gollapudi R, Goldberg R, Rapeport K, Haaser S, Topol S, Knowlton S, Bethel K, Kuhn P, Wood M, Carragher B, Schork NJ, Jiang J, Rao C, Connelly M, Fowler VM, Topol EJ. Characterization of circulating endothelial cells in acute myocardial infarction. Sci Transl Med. 2012 Mar 21;4(126):126ra33. doi: 10.1126/scitranslmed.3003451.
- Background: PubMed 22315715Aird WC. Endothelial cell heterogeneity. Cold Spring Harb Perspect Med. 2012 Jan;2(1):a006429. doi: 10.1101/cshperspect.a006429.
- Individual participant dataSharing planned: Yes. This project will reveal the first atlas (open acces) of circulating endothelial cells in human at both healthy and diabetes conditions.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Niels Jessen, Professor principal investigator
Steno Diabetes Center Aarhus, Aarhus Universitet Hospital
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Diabetes and Diabetes Mellitus, run by Aarhus University Hospital at 1 site in Denmark.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 3 inclusion and 2 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 20 Aug 2024. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Denmark. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Genome wide expression profile of circulating cells in blood samples from each cohort group (time frame: The PBMCs (including circulating endothelial cells) will be freshly isolated from the whole blood sampel, and frozen until all samples are collected (8 months). Afterwards flow cytometry and scRNA-seq analysis will be performed on all samples (2 months).). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT05169502, which also lists 4 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 20 Aug 2024.