Principles of reproducible metabolite profiling of enriched lymphocytes in tumors and ascites from human ovarian cancer.

Autor: Kilgour MK; Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada.; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., MacPherson S; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., Zacharias LG; Children's Research Institute, UT Southwestern Medical Center, Dallas, TX, USA., LeBlanc J; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada.; Biobanking and Biospecimen Research Services, Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., Babinszky S; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada.; Biobanking and Biospecimen Research Services, Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., Kowalchuk G; Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada.; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., Parks S; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada.; Biology Department, School of Arts & Science, Camosun College, Victoria, British Columbia, Canada., Sheldon RD; Department of Metabolism and Nutritional Programming, Van Andel Institute, Grand Rapids, MI, USA.; Metabolomics and Bioenergetics Core Facility, Van Andel Institute, Grand Rapids, MI, USA., Jones RG; Department of Metabolism and Nutritional Programming, Van Andel Institute, Grand Rapids, MI, USA.; Metabolomics and Bioenergetics Core Facility, Van Andel Institute, Grand Rapids, MI, USA., DeBerardinis RJ; Children's Research Institute, UT Southwestern Medical Center, Dallas, TX, USA.; Howard Hughes Medical Institute, UT Southwestern Medical Center, Dallas, TX, USA., Hamilton PT; Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada.; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., Watson PH; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada.; Biobanking and Biospecimen Research Services, Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada., Lum JJ; Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada. jjlum@bccancer.bc.ca.; Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, British Columbia, Canada. jjlum@bccancer.bc.ca.
Jazyk: angličtina
Zdroj: Nature protocols [Nat Protoc] 2022 Nov; Vol. 17 (11), pp. 2668-2698. Date of Electronic Publication: 2022 Aug 19.
DOI: 10.1038/s41596-022-00729-z
Abstrakt: Identifying metabolites and delineating their immune-regulatory contribution in the tumor microenvironment is an area of intense study. Interrogating metabolites and metabolic networks among immune cell subsets and host cells from resected tissues and fluids of human patients presents a major challenge, owing to the specialized handling of samples for downstream metabolomics. To address this, we first outline the importance of collaborating with a biobank for coordinating and streamlining workflow for point of care, sample collection, processing and cryopreservation. After specimen collection, we describe our 60-min rapid bead-based cellular enrichment method that supports metabolite analysis between T cells and tumor cells by mass spectrometry. We also describe how the metabolic data can be complemented with metabolic profiling by flow cytometry. This protocol can serve as a foundation for interrogating the metabolism of cell subsets from primary human ovarian cancer.
(© 2022. Springer Nature Limited.)
Databáze: MEDLINE
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