|
| Status |
Public on Mar 01, 2023 |
| Title |
B16F10_IL2-IL12_TwoDose_2d_C |
| Sample type |
RNA |
| |
|
| Source name |
Tumor
|
| Organism |
Mus musculus |
| Characteristics |
tumor model: B16F10
treatment: Intratumoral IL-2 and IL-12
cohort: Cohort 3
|
| Treatment protocol |
C57BL/6 mice bearing B16F10 tumors (day 6, ~30mm^2) were randomly assigned into three treatment cohorts to receive intratumoral collagen-binding cytokines IL-2 and IL-12 two days prior to tumor collection (cohort 1; n=3), eight days prior to collection (cohort 2; n=3), or two doses of cytokines (day 0 and day 6) with tumor collection two days after the second dose (cohort 3; n=3). Three mice also had tumors treated with saline as controls, with tumors removed two days after injection.
|
| Growth protocol |
B16F10 cells (ATCC) were maintained in 5% CO2 at 37C and tested negative for mycoplasma and rodent pathogens (IMPACT I test, IDEXX) prior to tumor inoculation of 1M cells into the right flank of C57BL/6 mice.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
RNA was extracted from FFPE tissue scrolls (3x10um) using Qiagen FFPE RNEasy kit following manufacturer's protocol. Samples were QC'd for concentration and fragment analysis on Bioanalyzer (Agilent) prior to hybridization to the nCounter panel probe set.
|
| Label |
n/a
|
| Label protocol |
n/a
|
| |
|
| Hybridization protocol |
Hybridization of RNA to the Mouse PanCancer Immune Profiler nCounter panel probe set (Nanostring) was performed according to manufacturer's instructions for 22 hours at 65C. Hybridized RNA was loaded into the analysis cartridge using Nanostring PrepStation.
|
| Scan protocol |
Scanning of analysis cartridge was performed on Nanostring Digital Analyzer under maximum resolution.
|
| Description |
Nanostring Mouse PanCancer Immune Profiling Panel (XT-CSO-MIP1-12)
|
| Data processing |
Nanostring nSolver software was used to process count data. Background thresholding was performed using the mean of 8 negative control probes. Normalization was performed in nSolver prior to analysis of differential gene expression and pathway analysis between cohorts.
|
| |
|
| Submission date |
Nov 28, 2022 |
| Last update date |
Mar 03, 2023 |
| Contact name |
K Dane Wittrup |
| E-mail(s) |
wittrup@mit.edu
|
| Phone |
6172534578
|
| Organization name |
Massachusetts Institute of Technology
|
| Department |
Koch Institute for Integrative Cancer Research
|
| Lab |
Wittrup Lab
|
| Street address |
500 Main St, 76-261
|
| City |
Cambridge |
| State/province |
Massachusetts |
| ZIP/Postal code |
02139 |
| Country |
USA |
| |
|
| Platform ID |
GPL30298 |
| Series (2) |
| GSE218913 |
Collagen-anchored interleukin-2 and interleukin-12 safely reprogram the tumor microenvironment in mouse melanomas |
| GSE218914 |
Collagen-anchored interleukin-2 and interleukin-12 safely reprogram the tumor microenvironment in soft tissue sarcomas |
|
