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Doshi, Kashish

University of British Columbia

Follicular Lymphoma (FL) is an indolent type of Non-Hodgkin’s Lymphoma comprising 20% of new lymphoma diagnosis with a risk of transforming into a more aggressive B-cell disease. Although recurrent mutations in FL are informative of transformation risk, biological predictors of transformation at diagnosis remain elusive. A previous study from our laboratory identified 2 phenotypic variation patterns, using the CyTOF, that were a significant predictor of transformation risk and implied biological diversity between FL patients. More recently, we assayed 8 samples from our previous cohort using multiomic single-cell techniques to investigate the cell-intrinsic sources of diversity. However, a multiomic analysis involves integration of the high dimensional single-cell sequencing datasets with the low dimensional CyTOF data. To address this issue, a bioinformatic pipeline was developed that integrated both low and high dimensional datasets using tools developed for single-cell integration. FastMNN and SingleR demonstrated efficacy for low dimensional dataset integration and annotation transfer. High dimensional integration was demonstrated using all tools identically. These findings offer a comprehensive framework to blend tools for analyzing low-parameter CyTOF and Flow Cytometry with high-parameter single-cell sequencing, enabling a more thorough examination of biology and disease progression. Additionally, our previous study identified two phenotypic clusters that comprised 50% of all malignant cells from 155 FL cases assayed using the CyTOF. One of these clusters, referred to as MC B, phenotypically similar to Naïve B-cells and expressed surface IgM. Given that 60% of FL cases exhibit recurrent mutations in chromatin-modifying genes (CMG), we hypothesized that these mutations could affect class switch recombination (CSR) in FL. To explore this hypothesis, a novel primary B-cell CSR model was established. My findings demonstrated that the addition of the ImmunoCult supplement enabled robust proliferation and CSR in primary Naïve B-cells. Moreover, successful transduction of Naïve B-cells was achieved using a Gibbon Ape Leukemia Virus (GaLV) based lentiviral vector. However, knockdown of CMG resulted in growth arrest of the transduced cells, highlighting the need for further refinement of the B-cell model before definitive conclusions can be drawn.

Text

2024-04-11

Attribution-NonCommercial-NoDerivatives 4.0 International

http://hdl.handle.net/2429/87764

Genome Science and Technology

University of British Columbia

UBCV

http://creativecommons.org/licenses/by-nc-nd/4.0/