Session: 651. Multiple Myeloma and Plasma Cell Dyscrasias: Basic and Translational: Poster I
Hematology Disease Topics & Pathways:
Research, Translational Research, Plasma Cell Disorders, Diseases, Immunology, Lymphoid Malignancies, Metabolism, Biological Processes, Technology and Procedures, Omics technologies
The tumor ecosystem in multiple myeloma (MM) involves complex interactions among various cell types, including clonal plasma cells (PCs) and immune cells. Previous research has indicated that the bone marrow (BM) microenvironment's metabolite and lipid profiles differ between MM patients and those with precursor conditions. This study aims to explore if differences in cell composition, metabolic phenotype of clonal PCs, and the spatial organization of various cell types are linked to the early versus late progression of high-risk smoldering MM (SMM) to MM.
Methods
We identified high-risk SMM patients using the IMWG 20-2-20 criteria, focusing on early progressors (EP), who progressed to MM within one year, and later progressors (LP), who progressed after more than four years. Using a high-dimensional single-cell spatial imaging platform, CODEX (CO-Detection by indEXing), we identified clonal PCs and immune cell types as well as their metabolic phenotypes at the single-cell level, in paired FFPE BM samples at SMM diagnosis and at the time of progression to MM. Our validated panel included markers such as CD3e, CD4, CD8, CD14, CD20, CD31, CD38, CD45, CD66a, CD68, CD138, HLA-DR, ASCT2, pNRF2, GLUT1, LDHA, ATPA5, CPT1A, and Citrate synthase. We employed single-cell segmentation, artifact cleanup, signal thresholding, and spatial analysis. Repeated measure models accommodated multiple regions of interest (ROIs) per patient and accounted for potential intrapatient correlation, while Poisson models estimated incidence rate ratios (RR) of cell types. Spatial statistical models evaluated cell type segregation and integration at the time of SMM diagnosis.
Results
Single cell events were acquired from 6 unique SMM patients who had paired FFPE BM samples from the time of their SMM diagnosis and at the time of their progression to MM. There were 3 EP SMM patients (Time to Progression (TTP):8, 8, 10 mos) and 3 LP SMM patients (TTP: 51, 58, 117 mos). The median number of single cell events identified per patient slide sample was 104,390 (Range: 24,357 – 314,877).
We identified 8 main cell types: PCs, CD4+ T-cells, CD8+ T-cells, B-cells, granulocytes, Monocyte/Macrophages and two groups of other immune cells. The PCs and the remainder of the non-PCs were further classified based on the expression of the following metabolic markers: ASCT2 (Glutamine importer), CPT1A (Fatty acid metabolism), GLUT1 (Glucose importer), LDHA (Glycolysis), Citrate synthase and ATPA5 (Oxidative Phosphorylation). There was a higher RR of granulocytes (RR 2.1, CI 1.6-2.8, p<0.001) in EP compared to LP samples at the time of SMM diagnosis, but a lower RR of CD4 T cells (RR 0.40, CI 0.3-0.6, p< 0.001). Focusing on metabolic phenotypes of the PCs, we observed higher incidence RR of most metabolic markers: ASCT2 (RR=1.77; p=0.032), ATPA5 (RR=1.25; p<0.001), CPT1a (RR=2.42; p=0.015), and LDHA (RR 2.13; p<0.001) in EP compared to LP samples at the time of SMM diagnosis. However, GLUT1 had a lower incidence RR in EP patients (RR=0.79; p<0.001). The same modeling for non-PCs at the time of SMM diagnosis yielded higher expression of LDHA (RR=1.98; p<0.001) and ASCT2 (RR=3.3; p=0.002).
Using the x and y coordinates and the corresponding phenotype classifications, we performed spatial statistical modeling based on spatial point processes to generate aggregated measures of how different types of cells of interest interacted. CD4 T cells were more isolated in LPs compared to EPs (p =0.046). Segregation indices for PCs and CD4 T cells indicated that they were more integrated/collocated in EPs. Granulocytes were more isolated in EPs although the difference was not as pronounced (p= 0.07). PCs were more integrated with granulocytes (p=0.06) and B cells (p=0.07) in LP patients. Further analyses of paired samples at the time of progression to MM will be reported at the time of the presentation.
Conclusion
This study is the first to provide whole-slide, single-cell, spatial proteomic characterization of clonal PCs and immune cells in the FFPE BMs of high-risk SMM patients with varying progression times to MM. Distinct metabolic pathway markers in clonal PCs and their spatial relationships with immune cell subsets are closely associated with the risk of progression to MM. These findings illuminate the complex interactions between the metabolic states of clonal PCs and their spatial orientation relative to immune cells in the pathogenesis of MM.
Disclosures: Soekojo: Smart immune: Honoraria; J&J: Honoraria; Sanofi: Honoraria; Antegene: Honoraria. Kumar: Merck: Research Funding; MedImmune/AstraZeneca: Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche: Research Funding; Takeda: Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen: Membership on an entity's Board of Directors or advisory committees, Research Funding; Oncopeptides: Other: Independent review committee participation; Novartis: Research Funding; Sanofi: Research Funding; Adaptive: Membership on an entity's Board of Directors or advisory committees, Research Funding; Abbvie: Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; KITE: Membership on an entity's Board of Directors or advisory committees, Research Funding. Cook: Geron Corp: Other: Held $600 Geron Stock for one week and sold without profit . Dispenzieri: Pfizer: Research Funding; HaemaloiX: Research Funding; Alexion: Consultancy, Research Funding; BMS: Consultancy, Research Funding; Alnylam: Research Funding; Janssen: Research Funding; Takeda: Consultancy, Research Funding. Fonseca: Patent for FISH in MM - ~$2000/year: Patents & Royalties: Patent for FISH in MM - ~$2000/year; AbbVie, Adaptive, Amgen, Apple, Bayer, BMS/Celgene, Gilead, GSK, Janssen, Kite, Karyopharm, Merck Sharp & Dohme, Juno Therapeutics, Takeda, Arduro Biotech, Oncotracker, Oncopeptides, Pharmacyclics, Pfizer, RA Capital, Regeneron, Sanofi: Consultancy; Antengene: Membership on an entity's Board of Directors or advisory committees; Celgene, Bristol Myers Squibb, Bayer, Amgen, Janssen, Kite, a Gilead company, Merck Sharp & Dohme, Juno Therapeutics, Takeda, AbbVie, Aduro Biotech, Sanofi, OncoTracker: Honoraria. Villasboas Bisneto: Genentech: Research Funding; Regeneron: Research Funding; Epizyme: Research Funding; Enterome: Research Funding; CRISPR: Research Funding; Aptose: Research Funding.
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