Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT05171998 |
| Other study ID # |
CT 253888 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
January 1, 2021 |
| Est. completion date |
June 1, 2021 |
Study information
| Verified date |
November 2021 |
| Source |
Cwm Taf University Health Board (NHS) |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
Emerging clinical details of the current SARS-CoV-2 pandemic have illustrated that there are
multiple clinical presentations and outcomes of this viral infection. People with an
infection have been reported to have a spectrum of disease from severe acute respiratory
distress requiring ventilation, to mild respiratory or gastrointestinal symptoms and
asymptomatic presentations. The SARS-CoV-2 pandemic has been accompanied with a substantial
increase in the number of individuals presenting with new onset type 1 diabetes [1]. Most
individuals presenting with type 1 diabetes since the start of the COVID-19 pandemic are
SARS-CoV-2 antibody positive. These findings suggest that SARS-CoV-2 infection can cause type
1 diabetes. Investigators have identified that many individuals presenting with type 1
diabetes since the start of the COVID-19 pandemic are SARS-CoV-2 positive by swab or blood
test. Researchers have also observed that T cells in patients who have had COVID recognise
some of the peptides in the pancreatic islet cells, which are responsible for production of
insulin. These findings suggest that SARS-CoV-2 infection may be associated with new onset of
type 1 diabetes.
The aim of this project is to understand the host immune response to infection with
SARS-CoV-2 over time in convalescent newly diagnosed patients with type 1 diabetes, including
acquired immune responses, gene expression profiling in peripheral blood and to identify host
genetic variants associated with disease progressions or severity. Participants will have
Type 1 diabetes and will have had a diagnosis of COVID-19 (confirmed by a positive
nasopharyngeal swab PCR test and/or SARS-CoV-2 antibody test) and have recovered from
COVID-19. Samples will be processed and analysed to explore the molecular mechanisms by which
SARS-CoV-2 infection might precipitate immune attack on insulin-producing cells resulting in
autoimmune diabetes.
Description:
AIMS AND OBJECTIVES The objective of this project is to understand the host immune response
to infection with SARS-CoV-2 over time in convalescent patients that develop type 1 diabetes
(T1D). Preliminary data show that diabetogenic T-cells in patients that developed T1D before
2019 can recognise SARS-CoV-2. If the reverse is true, it would provide a plausible mechanism
for the development of T1D following SARS-CoV-2 infection. We would like to ascertain why
some people develop T1D after SARS-CoV-2 infection.
Primary objectives are to determine whether individuals that develop T1D following SARS-CoV-2
infection bear the HLA A*0201 and HLA*2402 genes known to be associated with T1D and to show
that SARS-CoV-2-specific killer T-cells in these individuals are capable of directly
destroying insulin-producing pancreatic beta cells. The ability of SARS-CoV-2-specific
T-cells to recognise diabetogenic T-cell epitopes will be examined using peptide-HLA
multimers. Cross-reactive T-cell receptors will be manufactured for biophysical and
structural analyses to enable an understanding of how SARS-CoV-2 might induce T1D.
Methodology will examine whether convalescent COVID-19 patients that develop T1D
predominantly express the human leukocyte antigens (HLAs) HLA A*02 and HLA*24 by staining a
small part of the sample with antibodies for these HLA. Investigators anticipate that there
could be a large enrichment for these disease risk HLA (>75% of patients expressing one of
these disease-risk alleles) [2]. T-cell library [3] and peptide-HLA multimer [4-6] approaches
will be used to isolate T-cells that recognise epitopes from SARS-CoV-2 via HLA A*02 and
HLA*24. The monoclonal T-cell populations generated will be tested for recognition of known
diabetogenic epitopes present on the surface of human pancreatic beta cells. Where such
T-cell cross-reactivity [7] is identified the team will sequence the complementarity
determining region of the cognate T-cell receptor so that it can be manufactured as a soluble
protein [8] for further biophysical and structural studies [9,10] aimed at understanding how
SARS-CoV-2 might induce T1D in a minority of individuals.
The study will be prospective, observational study. Participants will be convalescent
patients with recent new diagnosis of type 1 diabetes (n=20). Eligible participants will be
identified and referred from Consultant Paediatric Endocrinologists at The Department of
Child Health at Cwm Taf Morgannwg University Health board, The Department of Child Health,
Cardiff and the Vale health board and Department of Child Health, St, Mary's Hospital,
Londonwho are all part of the clinical research study team and direct clinical care team. All
participants and their parents /guardians will receive a participant information sheet and
have time and privacy to read and understand the content. A member of the study team trained
in Good Clinical Practice will be available to provide support and answer any questions.
Informed consent will then be gained in writing on a consent form, or in the case of
paediatric participants on an assent form and a Parent/Guardian consent form. Participants
will be issued with a unique study number.
A sample of blood will be taken by a trained phlebotomist, 50mls (or less in children
according to the participant's weight). The sample will be anonymised and issued with the
unique study number. We will require peripheral blood mononuclear cells (PBMC) from of blood
on the day of recruitment. Anonymised blood samples will be transferred for T cell research
and Cwm Taf University Health board laboratory for serology tests (Roche SARS-CoV-2 IgM and
IgG assay at a UKAS accredited laboratory) with appropriate UN3373 packaging. Samples will be
used for immunological, virological and host genetic assays. Bloods will be processed to
isolate serum and PBMC, including host RNA and DNA. T-cell libraries will be established
using the PBMC. T-cell clones generated will be tested for cross-reactivity with established
diabetogenic T-cell epitopes. The T-cell receptors (TCRs) will be manufactured from
cross-reactive T-cells for further study. PBMC samples will also be stained with cognate
peptide-HLA multimers using flow cytometry. This will generate information about their
phenotype and activation status.
A Lay Description:
The blood will be separated into constituent parts using a thick density gradient solution
and a centrifuge. The white blood cell layer, or PBMCs, is isolated from other parts of the
blood and retained. All other parts are discarded in a bleach solution PBMCs are further
processed to isolate the T-cells, these cells can then go down one of five routes:
1. The cells are treated to extract the DNA/RNA, and the DNA/RNA is analysed to gain
information about the T-cell receptors and other associated molecules. The cells are
killed during DNA/RNA extraction.
2. The cells will undergo genetic editing to provide information about the processes
involved in normal T-cell function. Cells will be cultured for 28 days post gene
editing, will undergo multiple rounds of division, and become non-relevant in terms of
the Human Tissue Authority (HTA) act 2004.
3. The cells will be used in an assay where they are exposed to overlapping bits of
SARS-CoV-2 proteins, they will then be grown in culture for 14 days after which time the
cells will have undergone division and will be no longer HTA relevant. Cells that
respond to SARS-CoV-2 proteins will be tested to see whether they have capacity to
destroy insulin-producing pancreatic beta cells.
4. Using an automated sorter and fluorescent markers, the cells are sorted and analysed.
Cells will die during this process which takes place on the day cells are processed in
the lab.
5. Cells will be used in plate-based assays to learn about the processes involved in T-cell
functions. Cells will be disposed after the assay which can take between 1 and 3 days
depending on type of assay used.
The date and time of the participants positive SARS-CoV-2 swab/blood test will be captured
and linked to the participant study record using the unique study number by a member of the
research team in the direct clinical care teams These details will be stored on a secure NHS
server and in the site file at each hospital site in a secure, lockable, fireproof container.
All participants will be given a participant information sheet to take away with them. This
data set will not be accessible to laboratory-based researchers at any of the connected
universities.
This methodology was designed following Patient and Public Involvement, which were conducted
remotely due to social distancing.
