Summary of Study ST002489
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001607. The data can be accessed directly via it's Project DOI: 10.21228/M8013C This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
Study ID | ST002489 |
Study Title | Simultaneous targeting of PD-1 and IL2Rβγ with radiation therapy to inhibit pancreatic cancer growth and metastasis |
Study Summary | Mice were orthotopically implanted with PK5L1940 cells then radiation therapy (8 Gy) administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. T-cells from spleen were obtained through sorting, cultured in the presence of 13C6 glucose for 6h, and glucose fluxes profiled via mass spectrometry-based metabolomics. |
Institute | University of Colorado Denver |
Last Name | Haines |
First Name | Julie |
Address | 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA |
julie.haines@cuanschutz.edu | |
Phone | 3037243339 |
Submit Date | 2023-02-21 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2023-03-10 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
Project ID: | PR001607 |
Project DOI: | doi: 10.21228/M8013C |
Project Title: | Simultaneous targeting of PD-1 and IL2Rβγ with radiation therapy to inhibit pancreatic cancer growth and metastasis |
Project Summary: | In pancreatic ductal adenocarcinoma (PDAC) patients, we show that response to radiation therapy (RT) is characterized by increased IL2Rβ and IL2Rγ and decreased ILR2α expression. The bispecific aPD1-IL2v is a PD-1-targeted IL-2 variant (IL2v) immunocytokine with engineered IL-2 cis-targeted to PD-1 and abolished IL2Rα binding, which enhances tumor-antigen specific T cell activation while reducing regulatory T cell (Treg) suppression. Using aPD1-IL2v in orthotopic PDAC KPC-driven tumor models, we show marked improvement in local and metastatic survival along with profound increase in tumor-infiltrating polyfunctional CD8+ T cell subsets with a transcriptionally and metabolically active phenotype, and preferential activation of antigen-specific CD8+ T cells. In combination with single dose RT, aPD1-IL2v treatment results in a robust, durable expansion of polyfunctional CD8+ T cells, T cell stemness, tumor-specific memory immune response, natural killer (NK) cell activation, and decreased Tregs. These data show that aPD1-IL2v leads to profound local and distant response in PDAC. |
Institute: | University of Colorado Denver |
Laboratory: | Lab of Angelo D'Alessandro in collaboration with lab of Sana Karam |
Last Name: | Haines |
First Name: | Julie |
Address: | 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA |
Email: | julie.haines@cuanschutz.edu |
Phone: | 3037243339 |
Subject:
Subject ID: | SU002579 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Species Group: | Mammals |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
mb_sample_id | local_sample_id | factor |
---|---|---|
SA248865 | AG-23-TC-15 | aCD25 |
SA248866 | AG-23-TC-14 | aCD25 |
SA248867 | AG-23-TC-13 | aCD25 |
SA248856 | AG-23-TC-9 | IL2v |
SA248857 | AG-23-TC-7 | IL2v |
SA248858 | AG-23-TC-8 | IL2v |
SA248859 | AG-23-TC-5 | RT |
SA248860 | AG-23-TC-4 | RT |
SA248861 | AG-23-TC-6 | RT |
SA248862 | AG-23-TC-11 | RT+IL2v |
SA248863 | AG-23-TC-12 | RT+IL2v |
SA248864 | AG-23-TC-10 | RT+IL2v |
SA248868 | AG-23-TC-2 | unx |
SA248869 | AG-23-TC-1 | unx |
SA248870 | AG-23-TC-3 | unx |
Showing results 1 to 15 of 15 |
Collection:
Collection ID: | CO002572 |
Collection Summary: | C57BL/6 mice were orthotopically implanted with PK5L1940 cells. 8 Gy RT was administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. Spleens and lymph nodes of C57BL/6 mice were harvested 17 days post implantation and homogenized to a single cell suspension by mashing through a 70 uM cells strainer. For spleen processing, and the erythrocytes were lysed with ACK (ammonium-chloride-potassium) lysis buffer for 5 min at RT. CD8 T cells were sorted with a CD8-negative selection Stemcell isolation kit following manufacturer instructions. |
Sample Type: | T-cells |
Treatment:
Treatment ID: | TR002591 |
Treatment Summary: | CD8+ T cells were cultured for 6h in the presence of 25 mM U-13C-glucose in glucose-free RPMI. Cells were washed in PBS at 1200RPM, 4degC for 10 minutes and pellets were flash frozen. |
Sample Preparation:
Sampleprep ID: | SP002585 |
Sampleprep Summary: | Metabolites from frozen T-cell pellets were extracted with cold 5:3:2 MeOH:acetonitrile:water at a concentration of 4e6 cells/ml. Samples were vortexed 30 min at 4 degrees C then supernatants clarified by centrifugation (10 min, 10,000 g, 4 degrees C) and transferred to autosampler vials. |
Processing Storage Conditions: | 4℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN004063 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Thermo Vanquish |
Column | Phenomenex Kinetex C18 2.1 x 150 mm, 1.7 um |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap |
Ion Mode | NEGATIVE |
Units | peak area |
Chromatography:
Chromatography ID: | CH003009 |
Chromatography Summary: | Negative C18 |
Instrument Name: | Thermo Vanquish |
Column Name: | Phenomenex Kinetex C18 2.1 x 150 mm, 1.7 um |
Column Temperature: | 45 |
Flow Gradient: | 0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min 100-0% B, 3-5 min hold at 0% B |
Flow Rate: | 450 uL/min |
Sample Injection: | 10 uL |
Solvent A: | 95% water 5% acetonitrile 1 mM ammonium acetate |
Solvent B: | 95% acetonitrile 5% water 1 mM ammonium acetate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003810 |
Analysis ID: | AN004063 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database. |
Ion Mode: | NEGATIVE |