|
| Status |
Public on May 04, 2023 |
| Title |
CAR_Untrated_Donor#3 |
| Sample type |
RNA |
| |
|
| Source name |
PBMCs from Healthy Donor
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: Untreated CAR T cells
|
| Treatment protocol |
Un-transduced (NT) and CAR.CD19 T-cells were activated with 0.5 μg/ml recombinant human CD19 Fc Chimera Protein for 16 hours in presence or absence of emapalumab (100mg/ml) before the analysis of the gene signature.
|
| Growth protocol |
PBMCs derived from buffy coats of HDs were isolated, activated, transduced with the retroviral CAR.CD19 construct and expanded in media conaining IL7 and IL 15 cytokines.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated from samples using the RNeasy mini kit (Qiagen) according to supplier’s instructions.
|
| Label |
NanoString
|
| Label protocol |
All sample preparations were performed according to the manufacturer’s instructions (NanoString Technologies).
|
| |
|
| Hybridization protocol |
500 nanograms of total RNA were hybridized overnight with nCounter Reporter (20 μL) probes in hybridization buffer and excess of nCounter Capture probes (5 μL) at 65 °C for over night. After overnight hybridization, the abundance of specific target molecules was then quantified using the nCounter digital analyzer.
|
| Scan protocol |
Individual fluorescent barcodes and target molecules present in each sample were recorded with a CCD camera by performing a high-density scan (600 fields of view).
|
| Description |
Expression of 780 genes associated to essential pathways of CAR-T cell biology
|
| Data processing |
Raw data processing, quality control, and normalization were performed using the nSolver™ 4.0 analysis software (NanoString nCounter Technologies, Seattle, WA). Background subtraction from raw transcript counts was performed through negative input controls. Normalization to 10 housekeeping genes and differential expression analysis were completed using the Advanced Analysis software plugin (version 2.0.115). For differential expression analysis, a p‐value of ≤0.05, were applied as cut-offs.
|
| |
|
| Submission date |
Apr 04, 2023 |
| Last update date |
May 04, 2023 |
| Contact name |
Marika Guercio |
| E-mail(s) |
marika.guercio@opbg.net
|
| Phone |
0039 3339390943
|
| Organization name |
Ospedale Pediatrico Bambino Gesù
|
| Department |
Oncohematology
|
| Street address |
Viale di San Paolo, 15
|
| City |
Rome |
| State/province |
RM |
| ZIP/Postal code |
00146 |
| Country |
Italy |
| |
|
| Platform ID |
GPL29417 |
| Series (2) |
| GSE228921 |
Neutralizing IFNγ improves safety without compromising efficacy of CAR-T cell therapy in B-cell malignancies I |
| GSE228924 |
Neutralizing IFNγ improves safety without compromising efficacy of CAR-T cell therapy in B-cell malignancies |
|
