Primary Immunodeficiency Diseases (PID)
Conditions
Keywords
JAK inhibitors, STAT1 gain-of-function, gene therapy
Brief summary
The investigators want to study the JAK-inhibitors and their impact on the immune system and evaluate the potential of a gene-therapeutic strategy
Detailed description
The investigators want to study ex vivo the effect of JAK-inhibitors on the transcriptional profile and immune cell landscape in patients with inborn errors of the JAK-STAT pathway and the ex vivo evaluation of the feasibility of a gene therapeutic approach for STAT1 GOF. Following aspects will be compared: * To study pSTAT, transcriptional profile and cytokine production on bulk and sorted peripheral blood cell populations following stimulation in the presence or absence of different jakinibs * To evaluate to what extent jakinibs can normalize the transcriptional in different cell types (or not and identify blind spots of this treatment strategy) * To evaluate ex vivo the impact of a gene therapeutic approach for STAT1 GOF.
Interventions
DIAGNOSTIC_TESTblood sampling
Blood/serum samples will be collected during routine clinical visits at the time of planned peripheral venous blood sampling. Samples will be processed and either used immediately (flow cytometry-based cell sorting, gDNA extraction, in vitro functional assays, primary cell culture or single-cell applications) or stored for later analysis.
Sponsors
Research Foundation - Flanders (Fonds Wetenschappelijk Onderzoek)
prof. dr. Rik Schrijvers
Study design
Allocation
NON_RANDOMIZED
Intervention model
PARALLEL
Primary purpose
TREATMENT
Masking
NONE
Intervention model description
This is a parallel assignment study with two arms: (1) patients diagnosed with \[rare disease\] who will receive \[describe treatment\] and have blood drawn at specific time points, and (2) healthy controls who will undergo matched blood sampling for comparative analysis. Healthy controls will not receive the intervention.
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
Yes
Inclusion criteria
* Cases (A): adult patients presenting with a genetically confirmed or highly suspected disorder leading to an exagerated JAK-STAT pathway. * Controls (B): participants eligible for inclusion in this study must fall in one of the following categories: * Healthy controls (without immune-mediated disease)
Exclusion criteria
* Children (\< 18 years at time of recruitment) * Persons unable or unwilling to give informed consent
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Change in STAT phosphorylation levels in peripheral blood mononuclear cells (PBMCs) after cytokine stimulation with and without JAK inhibitor exposure | From time of inclusion to 24 months | Quantification via Median fluorescence intensity (MFI) of phosphorylated STAT1, STAT3, and STAT5 using multiparameter flow cytometry in bulk PBMCs and sorted T cells, B cells, NK cells, and monocyte subsets after standardized cytokine stimulation (e.g., IFNα, IFNγ, IL-6, IL-2) with or without JAK inhibitor exposure. Outcomes reported as fold-change relative to baseline. |
| Change in transcriptional profiles of immune cell subsets during JAK inhibitor treatment | From time of inclusion to 24 months | Differential gene expression assessed by single-cell RNA sequencing of PBMCs. Outcome is reported as the number of differentially expressed genes (adjusted p\<0.05) at different sampling timepoints (n=3) |
| Impact of ex vivo gene therapeutic correction in STAT1 gain-of-function patient-derived PBMCs | 24 months from inclusion | On-target editing efficiency measured as percentage of corrected alleles by targeted sequencing. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Transcriptional correction of our ex vivo gene therapeutic approach | 24 months | Transcriptional correction measured by single-cell RNA sequencing (fold-change of interferon pathway gene expression). |
| Mutation-specific differences in response to JAK inhibitor treatment | From time of inclusion to 24 months | Comparison of cytokine-induced phosphorylation (MFI of pSTAT1, pSTAT3, pSTAT5) and transcriptional responses (differential expression and pathway enrichment) between patients harboring distinct STAT1 gain-of-function variants. Outcomes reported as half-inhibitory concentration IC50 and area under the curve AUC in comparison to baseline |
| Functional differentiation capacity of gene therapy-corrected cells | 24 months | Functional differentiation capacity of corrected cells assessed by flow cytometry (percentages of T, B, NK, and monocyte subsets). |
| Impact of our gene therapeutic approach on cell viability | From time of inclusion to 24 months | Cell viability will be measured with MTT-assay |
| Off target events in our ex vivo gene therapeutic approach | 24 months | Off-target events detected by GUIDE-seq and confirmed by amplicon sequencing. |
Countries
Belgium
Contacts
Primary ContactRik Schrijvers, MD, PhD
Outcome results
None listed