000042699 001__ 42699
000042699 005__ 20240228123424.0
000042699 0247_ $$2doi$$a10.6083/bpxhc42699
000042699 037__ $$aETD
000042699 041__ $$aeng
000042699 245__ $$aBloodlines matter: leukemia associated mutation FLT3-ITD disrupts dendritic cell homeostasis and leads to altered T cell phenotypes
000042699 260__ $$bOregon Health and Science University
000042699 269__ $$a2024-02-24
000042699 336__ $$aDissertation
000042699 502__ $$bPh.D.
000042699 502__ $$gMolecular Microbiology & Immunology
000042699 520__ $$aDendritic cells (DC) are mediators between innate and adaptive immune responses to pathogens and tumors. DC development is determined by signaling through the receptor tyrosine kinase Fms-like tyrosine kinase 3 (FLT3) in bone marrow myeloid progenitors. Recently the naming conventions for DC phenotypes have been updated to distinguish between “Conventional” DCs (cDCs) and plasmacytoid DCs (pDCs). Activating mutations of FLT3, including Internal Tandem Duplication (FLT3-ITD), are associated with poor prognosis for acute myeloid leukemia (AML) patients. Having a shared myeloid lineage it can be difficult to distinguish bone fide DCs from AML tumor cells. To date, there is little information on the effects of FLT3-ITD in DC biology. To further elucidate this relationship we utilized CITE-seq technology in combination with flow cytometry and multiplex immunoassays to measure changes to DCs in human and mouse tissues. We examined the cDC phenotype and frequency in bone marrow aspirates from patients with AML to understand the changes to cDCs associated with FLT3-ITD. When compared to healthy donor (HD) we found that a subset of FLT3-ITD+ AML patient samples have overrepresented populations of cDCs and disrupted phenotypes. Using a mouse model of FLT3-ITD+ AML, we found that cDCs were increased in percentage and number compared to control wild-type (WT) mice. Single cell RNA-seq identified FLT3-ITD+ cDCs as skewed towards a cDC2 T-bet- phenotype, previously shown to promote Th17 T cells. We assessed the phenotypes of CD4+ T cells in the AML mice and found significant enrichment of both Treg and Th17 CD4+ T cells in the bone marrow and spleen compartments. Ex vivo stimulation of CD4+ T cells also showed increased Th17 phenotype in AML mice. Moreover, co-culture of AML mouse-derived DCs and naïve OT-II cells preferentially skewed T cells into a Th17 phenotype. Together, our data suggests that FLT3-ITD+ leukemia-associated cDCs polarize CD4+ T cells into Th17 subsets, a population that has been shown to be negatively associated with survival in solid tumor contexts. This illustrates the complex tumor microenvironment of AML and highlights the need for further investigation into the effects of FLT3-ITD mutations on DC phenotypes and their downstream effects on Th polarization.
000042699 540__ $$fCC BY-NC
000042699 542__ $$fIn copyright - single owner
000042699 650__ $$aAntigen-Presenting Cells$$014881
000042699 650__ $$aAllergy and Immunology$$014456
000042699 650__ $$aSingle-Cell Gene Expression Analysis$$013872
000042699 650__ $$aDendritic Cells$$017468
000042699 650__ $$aCytokines$$028964
000042699 6531_ $$aAML
000042699 6531_ $$aSingle Cell
000042699 6531_ $$aCD4 T cell
000042699 6531_ $$aCITE-seq
000042699 6531_ $$aRNAseq
000042699 6531_ $$aTh17
000042699 6531_ $$aCancer
000042699 6531_ $$acDCs
000042699 691__ $$aSchool of Medicine$$041369
000042699 692__ $$aDepartment of Molecular Microbiology and Immunology$$041429
000042699 7001_ $$aFlynn, Patrick$$uOregon Health and Science University$$041354$$10000-0003-4174-5562
000042699 8564_ $$yPAF 2024 PhD Thesis$$94a9b19a4-4eba-4830-99e1-fdd577da3ea3$$s8856571$$uhttps://digitalcollections.ohsu.edu/record/42699/files/Flynn.Patrick.2024.pdf
000042699 909CO $$ooai:digitalcollections.ohsu.edu:42699$$pstudent-work
000042699 980__ $$aTheses and Dissertations
000042699 981__ $$aPublished$$b2024-02-28