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

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

PROVIDER: PRJNA993275 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25224287.fastq.gz	Fastqsanger.gz
	SRR25224288.fastq.gz	Fastqsanger.gz
	SRR25224289.fastq.gz	Fastqsanger.gz
	SRR25224290.fastq.gz	Fastqsanger.gz
	SRR25224291.fastq.gz	Fastqsanger.gz

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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.

2024-01-03 | GSE236995 | GEO

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-TAMs are resistant to tumoricidal transformation by INFγ

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

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

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

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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.

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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.

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

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

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Gene expression profiles of tumor-associated macrophages (TAMs) overexpressing miR-511-3p

Project description:Hematopoietic stem/progenitor cells (HS/PCs) were transduced with lentiviral vectors overexpressing OFP and either miR-511-3p or a control, mutated miRNA sequence (miR-511-3p-mut). The transduced HS/PCs were then transplanted in recipient C57BL/6 mice. Tumors (Lewis lung carcinomas, LLC) were injected s.c. 4 weeks after HS/PC transplant. Lentiviral vector-transduced (OFP+), tumor-associated macrophages (TAMs) were isolated 4 weeks after LLC injection by fluorescence-activated cell sorting. Gene expression profiles of TAMs overexpressing either miR-511-3p or miR-511-3p-mut were obtained from 3 independent biological samples/each. Gene expression profiles of miRNA-overexpressing TAMs were then compared with the gene expression profiles of wild-type TAMs isolated from LLCs grown in nontransplanted C57BL/6 mice; the latter TAMs were subfractioned into MRC1+CD11c(low) or CD11c+MRC1(low) subsets before RNA isolation and analysis. Comparison of gene expression profiles of TAMs revealed that miR-511-3p overexpression tunes down the expression of multiple genes that are classically upregulated in protumoral MRC1+CD11c(low) TAMs. TAMs were isolated from Lewis lung carcinomas grown s.c. in C57BL/6 mice.

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