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Mixed cell-type spheroids endorse the generation and maintenance of heme-TAMs in an experimental tumor microenvironment

ABSTRACT: Mixed cell-type spheroids endorse the generation and maintenance of heme-TAMs in an experimental tumor microenvironment

PROVIDER: PRJNA991594 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25146892.fastq.gz	Fastqsanger.gz
	SRR25146893.fastq.gz	Fastqsanger.gz
	SRR25146894.fastq.gz	Fastqsanger.gz
	SRR25146895.fastq.gz	Fastqsanger.gz
	SRR25146896.fastq.gz	Fastqsanger.gz

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Mixed cell-type spheroids endorse the generation and maintenance of heme-TAMs in an experimental tumor microenvironment

Project description:To more accurately mimic the tumor microenvironment, we established a three-dimensional culture of MC38 cancer cells mixed with heme-treated BMDMs in microwell plates. This resulted in tumor spheroids with uniformly interspersed macrophages. We profiled the transcriptome of macrophages within these spheroids using scRNA-seq 24 hours after spheroid formation. To test the robustness of the heme effects in this model, we conducted a multiplexed experiment evaluating heme alone and in combination with the classical M1-polarization stimuli INFγ and LPS.

2024-01-03 | GSE236576 | GEO

Mixed cell-type spheroids endorse the generation and maintenance of heme-TAMs in an experimental tumor microenvironment [joint analysis]

Project description:Within the tumor microenvironment, heme exposure drives the transformation of macrophages into Arg1high Spp1high heme-TAMs with an enhanced matrix remodeling program and suppressed immunity, resisting inflammatory rewiring by INFγ and LPS.

2024-01-15 | GSE237257 | GEO

Heme-TAMs prevent cancer cell apoptosis and drive growth, invasiveness, and metastasis

Project description:To study the impact of heme-TAMs on the growth, phenotype, and function of cancer cells, we cultured GFP-MC38 cells either alone or mixed with heme-treated macrophages in microwell plates, enabling us to collect large numbers of uniform spheroids for scRNA-seq. We observed that after four days, the size of GFP-MC38 spheroids regressed, and the tumor cells underwent apoptosis and decayed metabolically. In contrast, the presence of heme-TAMs within spheroids prevented apoptosis, supporting the extended growth of metabolically viable spheroids beyond 10 days of culture.

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Heme-TAMs are resistant to tumoricidal transformation by INFγ

Project description:Inflammatory macrophage activation is a critical component of the antitumor immune response and an emerging therapeutic concept. To evaluate the impact of heme exposure on antitumoral macrophage activity, we grew spheroids of mixed GFP-MC38 and BMDMs that were either untreated or pretreated with heme, INFγ, or heme + INFγ, and characterized them by scRNA-seq.

2024-01-03 | GSE236996 | GEO

Heme-TAMs are resistant to tumoricidal transformation by INFγ

Project description:Heme-TAMs are resistant to tumoricidal transformation by INFγ

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Heme-TAMs prevent cancer cell apoptosis and drive growth, invasiveness, and metastasis

Project description:Heme-TAMs prevent cancer cell apoptosis and drive growth, invasiveness, and metastasis

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Heme-signaling progresses via NRF2 to educate heme-TAM transformation, tumor cell growth, and invasiveness

Project description:Heme-activation of NRF2 is a strong anti-inflammatory signal in macrophages, and analyses in this study indicated that the expressed transcriptome in heme-TAMs was consistently enriched for NRF2 target genes. We have therefore delineated the role of NRF2 in a series of experiments with Nrf2 knockout BMDMs, leading to a locked NRF2-inactive state, and macrophages with a knockout of the cytoplasmic NRF2 capture protein KEAP1, leading to a locked NRF2-active state, irrespective of the presence or absence of heme. To demonstrate that activated NRF2 is sufficient to drive heme-TAM transformation, we analyzed Keap1 knockout macrophages.

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Heme exposure shifts shifts myeloid differentiation in bone marrow derived mixed cell cultures.

Project description:Heme is an erythrocyte-derived toxin that drives disease progression in hemolytic anemias. During hemolysis, specialized bone marrow-derived macrophages with a high heme-metabolism capacity orchestrate disease adaptation by removing damaged erythrocytes and heme-protein complexes from the blood and supporting iron recycling for erythropoiesis. Here, we performed single-cell RNA sequencing with RNA velocity analysis of GM-CSF-supplemented mouse bone marrow cultures to assess myeloid differentiation under heme stress. We found that heme-activated NRF2 signaling shifted the differentiation trajectories of cells towards antioxidant, iron-recycling macrophages at the expense of dendritic cells, as these cells were selectively deficient in heme-exposed bone marrow cultures. Heme eliminated the capacity of GM-CSF-supplemented bone marrow cultures to activate antigen-specific T cells. The generation of functionally competent dendritic cells was restored by NRF2 loss. The heme-induced phenotype was reproduced in hemolytic mice with sickle cell disease and spherocytosis and associated with reduced dendritic cell functions in the spleen. Our data provide a novel mechanistic underpinning how hemolytic stress may provoke hyposplenism-related secondary immunodeficiency, which is a critical determinant of mortality in patients with genetic hemolytic anemias.

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Effects of Id1 depletion in TAMs on the genes expression of MC38 cells.

Project description:We report the application of high-throughput mRNA sequencing to explore the effect of Id1 depletion in TAMs on gene expression of MC38 cells. Different groups of TAMs were isolated from the subcutaneous tumors in Id1f/f or myeloid cell lineage speciﬁc Id1 deficient mice (Id1-Lyz-KO) . MC38 cells were then mixed with two groups of TAMs and be implanted in C57BL/6J mice. CD45- cells were then isolated from the subcutaneous tumors described above to do the transcriptome sequencing.

2023-10-15 | GSE188572 | GEO

Heme exposure shifts shifts myeloid differentiation in bone marrow derived mixed cell cultures.

Project description:Heme exposure shifts shifts myeloid differentiation in bone marrow derived mixed cell cultures.

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