Macrophage-Released Pyrimidines Inhibit Gemcitabine Therapy in Pancreatic Cancer

Halbrook, Christopher J.; Pontious, Corbin; Kovalenko, Ilya; Lapienyte, Laura; Dreyer, Stephan; Lee, Ho‐Joon; Thurston, Galloway; Zhang, Yaqing; Lazarus, Jenny; Sajjakulnukit, Peter; Hong, Hanna; Kremer, Daniel M.; Nelson, Barbara Scott; Kemp, Samantha B.; Zhang, Li; Chang, David K.; Biankin, Andrew V.; Shi, Jiaqi; Frankel, Timothy L.; Crawford, Howard C.; Morton, Jennifer P.; Magliano, Marina Pasca di; Lyssiotis, Costas A.

doi:10.1016/j.cmet.2019.02.001

Cited by 328 publications

(243 citation statements)

References 43 publications

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“…Next, RAW264.7 macrophages co-cultured in 3D with PyVMT breast cancer displayed changes in redox ratio consistent with metabolic changes from tumor stimulation in previous studies ( Fig 4A&C) 90 . Differences in redox ratio and fluorescence lifetime measurements were observed between RAW264.7 macrophages in cytokine-stimulated 2D cultures (Fig 2B-D) and tumor-stimulated 3D cultures ( Fig 4C-E), consistent with reported differences in intracellular metabolite concentration and metabolic flux based on stimulation condition (e.g., cytokines, tumor conditioned media) and culture in 2D vs. 3D 91,92 . Additionally, 3D co-cultured RAW264s exhibited heterogeneous cell-level autofluorescence and pooled gene expression ( Fig 4B, F-H).…”

Section: Discussionsupporting

confidence: 84%

Autofluorescence imaging of 3D tumor-macrophage microscale cultures resolves spatial and temporal dynamics of macrophage metabolism

Heaster

Humayun

et al. 2020

Preprint

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Macrophages within the tumor microenvironment (TME) exhibit a spectrum of pro-tumor and antitumor functions, yet it is unclear how the TME regulates this macrophage heterogeneity. Standard methods to measure macrophage heterogeneity require destructive processing, limiting spatiotemporal studies of function within the live, intact 3D TME. Here, we demonstrate twophoton autofluorescence imaging of NAD(P)H and FAD to non-destructively resolve spatiotemporal metabolic heterogeneity of individual macrophages within 3D microscale TME models. Fluorescence lifetimes and intensities of NAD(P)H and FAD were acquired at 24, 48, and 72 hours post-stimulation for mouse macrophages (RAW 264.7) stimulated with IFN-γ or IL-4 plus IL-13 in 2D culture, validating that autofluorescence measurements capture known metabolic phenotypes. To quantify metabolic dynamics of macrophages within the TME, mouse macrophages or human monocytes (RAW264.7 or THP-1) were cultured alone or with breast cancer cells (mouse PyVMT or primary human IDC) in 3D microfluidic platforms. Human monocytes and mouse macrophages in tumor co-cultures exhibited significantly different FAD mean lifetimes and greater migration than mono-cultures at 24, 48, and 72 hours post-seeding. In co-cultures with primary human cancer cells, actively-migrating monocyte-derived macrophages had greater redox ratios (NAD(P)H/FAD intensity) compared to passively-migrating monocytes at 24 and 48 hours post-seeding, reflecting metabolic heterogeneity in this subpopulation of monocytes. Genetic analyses further confirmed this metabolic heterogeneity. These results establish label-free autofluorescence imaging to quantify dynamic metabolism, polarization, and migration of macrophages at single-cell resolution within 3D microscale models. This combined culture and imaging system provides unique insights into spatiotemporal tumorimmune crosstalk within the 3D TME.

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Section: Discussionsupporting

confidence: 84%

Autofluorescence imaging of 3D tumor-macrophage microscale cultures resolves spatial and temporal dynamics of macrophage metabolism

Heaster

Humayun

et al. 2020

Preprint

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“…Certain chemotherapeutic agents, for example doxorubicin, enhance TAM accumulation in the tumour microenvironment, ultimately attenuating their cytotoxic effects . Although the molecular underpinnings are still enigmatic, recent work by Lyssiotis’ group showed that in murine models of pancreatic ductal adenocarcinoma, pyrimidine species released by TAMs inhibit gemcitabine via functional interference with drug uptake and metabolism . This suggests that TAM metabolism is interwoven with that of cancer cells with the potential to modulate the therapeutic response of solid tumours.…”

Section: The Role Of the Innate Immune System In Ccamentioning

confidence: 99%

The tumour microenvironment and immune milieu of cholangiocarcinoma

Fabris

Perugorria

Mertens

et al. 2019

Liver International

139

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Tumour microenvironment is a complex, multicellular functional compartment that, particularly when assembled as an abundant desmoplastic reaction, may profoundly affect the proliferative and invasive abilities of epithelial cancer cells. Tumour microenvironment comprises not only stromal cells, mainly cancer‐associated fibroblasts, but also immune cells of both the innate and adaptive system (tumour‐associated macrophages, neutrophils, natural killer cells, and T and B lymphocytes), and endothelial cells. This results in an intricate web of mutual communications regulated by an extensively remodelled extracellular matrix, where the tumour cells are centrally engaged. In this regard, cholangiocarcinoma, in particular the intrahepatic variant, has become the focus of mounting interest in the last years, largely because of the lack of effective therapies despite its rising incidence and high mortality rates worldwide. On the other hand, recent studies in pancreatic cancer, which similarly to cholangiocarcinoma, is highly desmoplastic, have argued against a tumour‐promoting function of the tumour microenvironment. In this review, we will discuss recent developments concerning the role of each cellular population and their multifaceted interplay with the malignant biliary epithelial counterpart. We ultimately hope to provide the working knowledge on how their manipulation may lead to a therapeutic gain in cholangiocarcinoma.

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“…In order to determine differences in the basal metabolic state between PDA and CRC cells, we used LC/MS-based metabolomics [21][22][23][24] to profile a panel of 3 PDA and 3 CRC parental cell lines in exponential growth phase. Analysis of statistically significant differences in the relative abundance of the steady state metabolite pools indicated that the PDA lines had more gluconodelta lactone-6 phosphate (GdL6P) and 6-phospho gluconate (6PG), metabolites in the oxidative arm of the PPP, and smaller metabolite pools of alanine and lactate ( Fig.…”

Section: Differential Metabolic Pathway Activity Between Pda and Crcmentioning

confidence: 99%

“…Thus, we set out to further interrogate the metabolic differences between GOT1 dependent and independent cells, and to determine differential central carbon utilization. To this end we performed isotope tracing metabolomics using either uniformly-labeled 13C (U-13 C) glucose (Glc) or glutamine (Gln) [22][23][24] in the parental PDA and CRC lines. Metabolites were collected from log phase cell lines grown overnight in labeled media, and fractional labeling patterns (Extended Fig.…”

Section: Differential Metabolic Pathway Activity Between Pda and Crcmentioning

confidence: 99%

Tissue of Origin Dictates GOT1 Dependence and Confers Synthetic Lethality to Radiotherapy

Nelson

Lin

Kremer

et al. 2019

Preprint

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Metabolic programs in cancer cells are influenced by genotype and the tissue of origin. We have previously shown that central carbon metabolism is rewired in pancreatic ductal adenocarcinoma (PDA) to support proliferation through a glutamate oxaloacetate transaminase 1 (GOT1)-dependent pathway. Here we tested if tissue type impacted GOT1 dependence by comparing PDA and colorectal cancer (CRC) cell lines and tumor models of similar genotype. We found CRC to be insensitive to GOT1 inhibition, contrasting markedly with PDA, which exhibit profound growth inhibition upon GOT1 knockdown. Utilizing a combination of metabolomics strategies and computational modeling, we found that GOT1 inhibition disrupted glycolysis, nucleotide metabolism, and redox homeostasis in PDA but not CRC. These insights were leveraged in PDA, where we demonstrate that radiotherapy potently enhanced the effect of GOT1 inhibition on tumor growth. Taken together, these results illustrate the role of tissue type in dictating metabolic dependencies and provide new insights for targeting metabolism to treat PDA.

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Macrophage-Released Pyrimidines Inhibit Gemcitabine Therapy in Pancreatic Cancer

Cited by 328 publications

References 43 publications

Autofluorescence imaging of 3D tumor-macrophage microscale cultures resolves spatial and temporal dynamics of macrophage metabolism

Autofluorescence imaging of 3D tumor-macrophage microscale cultures resolves spatial and temporal dynamics of macrophage metabolism

The tumour microenvironment and immune milieu of cholangiocarcinoma

Tissue of Origin Dictates GOT1 Dependence and Confers Synthetic Lethality to Radiotherapy

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