Seminar Talks—Five speakers will share insights, research, and results.

Luncheon—Mingle with other researchers from the surrounding area.

Step-Right-Up Q&A Table—Discuss your current projects and future ideas with an application scientist. Brand new to Mass Cytometry or Imaging Mass Cytometry? This is the perfect place to start.

Program

Continental Breakfast

cranberry scones, miniature banana muffins, lemon blueberry tea bread

bottled water, regular and decaf Starbucks coffee, assorted Tazo tea

Executive Welcome:

Jonathan Day, VP Commercial Operations, Fluidigm Corporation

Speaker 1: Cynthia Guidos, PhD

Senior Scientist, Developmental and Stem Cell Biology Program, Hospital for Sick Children Research Institute in Toronto, ON, Canada; Professor of Immunology, Faculty of Medicine, University of Toronto

“High Dimensional Characterization of Immune Cells in Normal and Diseased Micro-environments “

Abstract : Dissecting disease-driving immune responses presents a major analytical challenge because they are generally mediated by concomitant effects of diverse immune cell lineages, which are highly dynamic and heterogeneous at both the individual and the population levels. Thus, highly dimensional single cell and systems-wide approaches are needed to comprehensively characterize the immune cells in health and disease. Helios mass cytometers can measure expression of up to 48 different markers per cell, making CyTOF the leading technology for discovering biomarkers of immune-mediated diseases. In her presentation, Dr. Guidos will discuss how she is using mass cytometry to deeply characterize complex cellular immune networks that underlie anti-cancer immune responses and autoimmune diseases. She will describe how her group uses clustering, machine learning and dimensionality reduction algorithms to automatically identify and visualize differences in subset distribution and functional states across samples in an unbiased fashion. Finally, Dr. Guidos will overview her use of Fluidigm’s Hyperion multi-plexed imaging mass cytometry platform to enable deep characterization of immune cells within normal and diseased tissue microenvironments.

Speaker 2: Emily Thrash, PhD

Scientist II , Center for Immuno-Oncology – Immune Assessment Laboratory, Dana-Farber Cancer Institute

“Application of high-throughput mass cytometry to phenotype adaptive and innate immune subsets for clinical trials”

Biomarker discovery in clinical research is currently limited by the inherent complexity of cancer and immune responses in patients, as well as a lack of validated multi-parameter immuno-phenotyping methodology for large-scale trials. With more measurable parameters than traditional flow cytometry, mass cytometry (CyTOF) is poised to identify outcome-associated cellular signatures, follow disease progression and predict therapeutic responses. However, in a clinical setting, with large sample numbers collected at different timepoints, reduction of technical variability is critical. Using healthy donor PBMCs, we present the design and validation of two CyTOF panels focused on adaptive or innate immune response. Reference sample spike-in decreases batch effects between experiments and we demonstrate high-throughput sample number capabilities. Here we present our workflow and development for robustly immune-phenotyping clinical samples with CyTOF.

Speaker 3: Andrew Larry Frelinger, PhD

Assistant Professor of Pediatrics, Harvard Medical School, Associate Director, Center for Platelet Research Studies,Research Associate in Medicine, Division of Hematology/Oncology, Dana-Farber/Boston Children’s Cancer and Blood Disorders Center Boston Children’s Hospital/Dana-Farber Cancer Institute

Lunch

Traditional Sandwich Buffet

assorted traditional sandwiches: roasted turkey, roast beef, smoked ham, tuna salad, chicken salad, tomato, basil, mozzarella

FLIK chips, hand fruit

house baked FLIK signature chocolate chip cookies

bottled water, soda

Speaker 4: Douglas Linn, PhD

Associate Principal Scientist, Merck & Co., Inc.

“Imaging mass cytometry (IMC) as a powerful tool for immunophenotyping human and mouse tissues”

Abstract: The recent advent of imaging mass cytometry (IMC) applies the technologies of CyTOF to tissue imaging, building upon key weaknesses of flow cytometry and multiplex immunohistochemistry to establish a powerful tool for examining cellular interactions within complex microenvironments like tumors. We set out to establish and validate IMC capabilities within MRL Boston by analyzing tumors and lymphoid tissue stained with panels of human or mouse markers. Tissue processing and staining protocols were optimized to achieve simultaneous staining of dozens of immune markers, thus identifying major lymphocyte and myeloid subsets. Unsupervised clustering algorithms allowed examination of expression at population levels or single cell resolution. Together these efforts have validated the use of IMC to profile tumor and lymphoid tissues using large panels of immune markers. The highly multiplexed results that can be obtained will serve an important complementary role in immunophenotyping to explore biology and understand mechanism of action of cancer immunotherapies.

Speaker 5: Nikhil Singh, MD, PhD

Instructor, Section of Nephrology, Department of Internal Medicinel, Cantley Lab, Yale University

“Highly multiplexed, spatially preserved analysis of human kidney injury with Imaging Mass Cytometry”

Abstract: The lack of therapies for acute tubular injury (ATI) derives largely from our incomplete understanding of the biology of the injured human kidney. Although renal biopsy is commonly used in the diagnosis of acute kidney injury, the small amount of tissue obtained has previously limited the ability to use patient samples for discovery purposes. Imaging mass cytometry (IMC) is a novel technology which permits concurrent analysis of >40 markers on a formalin-fixed tissue section, ideal for scarce and archival samples. We have developed a highly reproducible methodology for IMC-based interrogation of the human kidney coupled to a machine-learning analysis pipeline. We have utilized this technique to perform comparative, quantitative, spatially preserved expression analysis of healthy and injured human tissue. With our combined imaging-analysis approach we have created a reference atlas for the adult human kidney, defining selected intrinsic and immune cell proportions and their spatial organization in 16 histopathologically normal samples. In injured kidneys with interstitial infiltrate, we have quantified both expected and novel changes to cellular proportions and tissue architecture. For a cohort of patients who underwent renal biopsy at two affiliated hospitals from 2011-2018 and whose biopsies showed ATI, we are adjudicating mechanism of injury by chart review and are interrogating banked tissue by IMC to identify immune drivers and cellular responses to injury, create a quantitative atlas of human ATI, and elucidate potential targets for therapeutic intervention. We hypothesize that unsupervised clustering of IMC data can be used to develop pathologic signatures corresponding to injury etiology and clinical outcome in ATI.