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Novel method to harvest viable cells allows for centralized RA research
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SAN DIEGO — A new method for harvesting viable cells from cryopreserved tissue samples could give researchers a model for collecting and analyzing samples from various study sites, allowing them to conduct more centralized research for rheumatoid arthritis, according to a presenter at the 2017 American College of Rheumatology Annual Meeting.
“The Accelerating Medicines Partnership program’s RA/SLE Network is an NIH partnership to try and analyze tissue samples in patients with rheumatoid arthritis in substantial detail,” Deepak Rao, MD, PhD, codirector of the Human Immunology Center at Brigham and Women’s Hospital, in Boston, and lead author of the study, said during the presentation. “The rationale behind this is that we recognize that although there has been substantial progress in the treatment of rheumatoid arthritis over many years, there still remains a substantial number of patients that fail to achieve sustained remission. Although we have therapies to treat rheumatoid arthritis, we have no strategies to help predict which patients will respond to which therapies.”
To acquire samples from patients with the rheumatoid arthritis, with a focus on synovial tissue samples, and to determine the dominant inflammatory pathways and cell types that characterize the disease, the researchers created a method to freeze the tissue intact soon after acquisition. The tissue was then stored, transported and analyzed at a single site. The researchers developed three detailed datasets on each sample, in a uniform way, so they could be compared.
According to the researchers, the preserved frozen tissue dissociated through these methods could be studied by multiple, high-dimensional analyses. Mass cytometry revealed diverse fibroblast phenotypes, clear separation of memory B cells from antibody-secreting cells and multiple phenotypes of activated CD4+ and CD8+ T cells, the researchers found. In addition, they developed a flow cytometric sorting strategy to collect fibroblasts, macrophages, T cells and B cells for bulk and single cell RNA-sequencing transcriptomics.
“This is interesting because, for one, it allows the acquisition of samples from many different sites, and the samples can then be preserved in a way that retains their integrity and allows them to be brought together at one site, where you can process them,” Rao said. “It’s advantageous because if one wants to use analyses like these, that are new, cutting-edge, state of the art analyses, they are not available at every site. So, you need to be able to bring those samples together to a single site, or limited number of sites, to analyze the samples.”
Reference:
Rao D. #1406. Presented at the American College of Rheumatology Annual Meeting, Nov. 3-8, 2017; San Diego.
Disclosure: Rao reports no relevant financial disclosures.