RESUMO
Biobanking of patient-derived specimens offers unique opportunities for retrospective testing that could potentially contribute to diagnosing and evaluating clinical conditions, advancing personalized medicine and translational biomedical discovery. In this protocol, we detail the collection, processing, and cryopreservation of peripheral blood, bone marrow, and lymph nodes from patients with hematological malignancies. This protocol can be used for multiomics to gain cellular and molecular insights into blood cancers and to test the therapeutic potential of compounds for translational biomedical research. For complete details on the use and execution of this protocol, please refer to Lim et al.1 and Rijal et al.2.
RESUMO
The innate immune response contributes to the development or attenuation of acute and chronic diseases, including cancer. Microbial DNA and mislocalized DNA from damaged host cells can activate different host responses that shape disease outcomes. Here, we show that mice and humans lacking a single allele of the DNA repair protein Ku70 had increased susceptibility to the development of intestinal cancer. Mechanistically, Ku70 translocates from the nucleus into the cytoplasm where it binds to cytosolic DNA and interacts with the GTPase Ras and the kinase Raf, forming a tripartite protein complex and docking at Rab5+Rab7+ early-late endosomes. This Ku70-Ras-Raf signalosome activates the MEK-ERK pathways, leading to impaired activation of cell cycle proteins Cdc25A and CDK1, reducing cell proliferation and tumorigenesis. We also identified the domains of Ku70, Ras, and Raf involved in activating the Ku70 signaling pathway. Therapeutics targeting components of the Ku70 signalosome could improve the treatment outcomes in cancer.
