Ignite Creation Date:
2024-05-06 @ 8:15 PM
Last Modification Date:
2024-10-26 @ 3:23 PM
Study NCT ID:
NCT06311123
Status:
RECRUITING
Last Update Posted:
2024-03-15
First Post:
2024-03-07
Brief Title:
Understanding Ozanimods MOA Via Mass Cytometry in Ulcerative Colitis
Sponsor:
University of California San Diego
Organization:
University of California San Diego
Study Overview
Official Title:
High Dimensional Mass Cytometry as a Tool to Understand Ozanimods Mechanism of Action MOA
Status:
RECRUITING
Status Verified Date:
2024-03
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
The goal of this observational study is to learn about the mechanism of action of ozanimod in patients with ulcerative colitis UC The main questions it aims to answer are
1 Prospectively assess the effects of ozanimod on the cellular composition of intestinal lamina propria and blood by deep immunophenotyping CyTOF of immune cell subsets prior and after the drugs administration
2 Determine whether changes in cell subsets observed via mass cytometry correlate with with clinical or histologic parameters of disease activity
Colonic biopsies and peripheral blood samples will be collected from patients with UC before and after the onset of ozanimod
Researchers will compare intestinal and peripheral leukocytes before and after the drugs administration
1 Prospectively assess the effects of ozanimod on the cellular composition of intestinal lamina propria and blood by deep immunophenotyping CyTOF of immune cell subsets prior and after the drugs administration
2 Determine whether changes in cell subsets observed via mass cytometry correlate with with clinical or histologic parameters of disease activity
Colonic biopsies and peripheral blood samples will be collected from patients with UC before and after the onset of ozanimod
Researchers will compare intestinal and peripheral leukocytes before and after the drugs administration
Detailed Description:
Study DesignMethodology
Several gastroenterologists at our center have agreed to assist with biopsy collection at our hospitals UCSD Singh Boland Patel VA Rivera-Nieves Dr Brian Behm who cares for most IBD patients at the University of Virginia has also agreed to assist with biopsyPBMC collection
During two visits T0 T24-32 we will collect biopsies endoscopies histology fecal and blood samples Table 1 prospectively from a total of 10 UC patients scheduled for colonoscopy before therapy with ozanimod and at 24-32 weeks post-treatment A sample size of 10 patients per condition should be sufficient to achieve significant differences using CITRUS based on required sample size determinations see below
The effects of the drug on cellular subsets will be assessed by comparing the two-time points 24-32 weeks post therapy with its pretreatment control T0 We will additionally compare the mass cytometry results to clinical parameters that are collected at our IBD center during every visit Table 1 The clinical samples and parameters from patients recruited at UVA will be sent de-identified through a secure mailbox managed by the UVA
Sample collection storage testing and analyses
1 After IV placement for conscious sedation 10 mL of blood will be drawn on EDTA-containing tubes and sent to the lab on ice for further processing Specimen transport is provided by staff from UCSD Center for Translational Research CTRI Upon arrival to the wet lab blood is spun plasma separated and leukocytes isolated by gradient centrifugation Cryopreservation media is added to the leukocyte fraction Plasma and leukocytes are stored at -80 C
2 Standard biopsies will also be obtained from ileum control effect of drug on uninvolved segment as well as in involved and uninvolved colonic segments and suspended in complete media for cell isolation Specimen transport is provided by staff from UCSD Center for Translational Research Upon arrival to the wet lab cryopreservation media is added to the biopsies and stored at -80 C
We have found significant batch to batch variation during pilot studies even controlled using veri-cells For that reason all celltissue processing and acquisition on the cytometer will all be performed together upon completion of collection
No study specific visits will be conducted for IBD patients All visits and sample acquisitions will be at standard of care visits No specific treatment related research interventions will be performed on IBD patients for this study We thus categorize the the study as non-interventional
Data Analysis
We have been analyzing CyTOF data now for several years as illustrated in the two published manuscripts by Tyler et al In addition we have worked with Amir El-Ad who runs the AstoLabe platform for high dimensional cytometery analyses To examine differences in population abundance and specific cell marker expression we have identified CITRUS as one of many appropriate algorithms to perform our analysis of IBD biopsies CITRUS identifies clusters of phenotypically similar cells in an unsupervised manner characterizes the behavior of identified clusters using biologically interpretable metrics and leverages regularized supervised learning algorithms to identify subset clusters whose behavior is predictive of the samples endpoint CITRUS strongly encourages the use of a minimum of 8 samples per group Thus for our current project we will obtain a minimum of 10 samples for UC before and after ozanimod therapy As per estimates based on our preliminary data this sample size will be sufficient to accurately identify even rare populations and to determine changes in abundance between cell subsets It is sensitive enough to assess differential expression of any marker in a three-way analysis Bioinformatics comparisons between the cell subsets present in ileum colon and PBMC at times 0 and 24-32 weeks for each patient will be performed for the principal endpoint of assessing the effect of the drug on relevant cell subsets
Most recently all of our data is stored in the Omiq platform which provides multiple algorithms for data analyses
For the univariable association of continuous independent variables with response either an unpaired t-test or a Wilcoxon two-sample test will be used while for categorical variables Fishers exact test will used All comparisons will be two-sided and performed at the 005 level of significance As these analyses are exploratory no adjustment will be made for multiple comparisons
Data Management Plan All data will be stored on password-protected computers at UCSD All hard files and raw data will be stored in locked cabinets on UCSD campus within the lab of Jesus Rivera-Nieves Patients will be given study identifiers as part of the IBD Biobank and no personal identifiers will be used The Data Management plan will follow that of the UCSD IBD Biobank which has been extensively reviewed by the IRB and Committee for Protection of Human Subjects at UCSD
At the UVA data will be collected by the local study coordinator and de-identified All data will be sent electronically using a secure mailbox This has been approved by the UVA IRB
Several gastroenterologists at our center have agreed to assist with biopsy collection at our hospitals UCSD Singh Boland Patel VA Rivera-Nieves Dr Brian Behm who cares for most IBD patients at the University of Virginia has also agreed to assist with biopsyPBMC collection
During two visits T0 T24-32 we will collect biopsies endoscopies histology fecal and blood samples Table 1 prospectively from a total of 10 UC patients scheduled for colonoscopy before therapy with ozanimod and at 24-32 weeks post-treatment A sample size of 10 patients per condition should be sufficient to achieve significant differences using CITRUS based on required sample size determinations see below
The effects of the drug on cellular subsets will be assessed by comparing the two-time points 24-32 weeks post therapy with its pretreatment control T0 We will additionally compare the mass cytometry results to clinical parameters that are collected at our IBD center during every visit Table 1 The clinical samples and parameters from patients recruited at UVA will be sent de-identified through a secure mailbox managed by the UVA
Sample collection storage testing and analyses
1 After IV placement for conscious sedation 10 mL of blood will be drawn on EDTA-containing tubes and sent to the lab on ice for further processing Specimen transport is provided by staff from UCSD Center for Translational Research CTRI Upon arrival to the wet lab blood is spun plasma separated and leukocytes isolated by gradient centrifugation Cryopreservation media is added to the leukocyte fraction Plasma and leukocytes are stored at -80 C
2 Standard biopsies will also be obtained from ileum control effect of drug on uninvolved segment as well as in involved and uninvolved colonic segments and suspended in complete media for cell isolation Specimen transport is provided by staff from UCSD Center for Translational Research Upon arrival to the wet lab cryopreservation media is added to the biopsies and stored at -80 C
We have found significant batch to batch variation during pilot studies even controlled using veri-cells For that reason all celltissue processing and acquisition on the cytometer will all be performed together upon completion of collection
No study specific visits will be conducted for IBD patients All visits and sample acquisitions will be at standard of care visits No specific treatment related research interventions will be performed on IBD patients for this study We thus categorize the the study as non-interventional
Data Analysis
We have been analyzing CyTOF data now for several years as illustrated in the two published manuscripts by Tyler et al In addition we have worked with Amir El-Ad who runs the AstoLabe platform for high dimensional cytometery analyses To examine differences in population abundance and specific cell marker expression we have identified CITRUS as one of many appropriate algorithms to perform our analysis of IBD biopsies CITRUS identifies clusters of phenotypically similar cells in an unsupervised manner characterizes the behavior of identified clusters using biologically interpretable metrics and leverages regularized supervised learning algorithms to identify subset clusters whose behavior is predictive of the samples endpoint CITRUS strongly encourages the use of a minimum of 8 samples per group Thus for our current project we will obtain a minimum of 10 samples for UC before and after ozanimod therapy As per estimates based on our preliminary data this sample size will be sufficient to accurately identify even rare populations and to determine changes in abundance between cell subsets It is sensitive enough to assess differential expression of any marker in a three-way analysis Bioinformatics comparisons between the cell subsets present in ileum colon and PBMC at times 0 and 24-32 weeks for each patient will be performed for the principal endpoint of assessing the effect of the drug on relevant cell subsets
Most recently all of our data is stored in the Omiq platform which provides multiple algorithms for data analyses
For the univariable association of continuous independent variables with response either an unpaired t-test or a Wilcoxon two-sample test will be used while for categorical variables Fishers exact test will used All comparisons will be two-sided and performed at the 005 level of significance As these analyses are exploratory no adjustment will be made for multiple comparisons
Data Management Plan All data will be stored on password-protected computers at UCSD All hard files and raw data will be stored in locked cabinets on UCSD campus within the lab of Jesus Rivera-Nieves Patients will be given study identifiers as part of the IBD Biobank and no personal identifiers will be used The Data Management plan will follow that of the UCSD IBD Biobank which has been extensively reviewed by the IRB and Committee for Protection of Human Subjects at UCSD
At the UVA data will be collected by the local study coordinator and de-identified All data will be sent electronically using a secure mailbox This has been approved by the UVA IRB
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None