DRAM1 increases the secretion of PKM2-enriched EVs from hepatocytes to promote macrophage activation and disease progression in ALD

Tan, Jie; Zhang, Jie; Wang, Mengke; Wang, Yifen; Dong, Mingjie; Ma, Xuefeng; Sun, Baoqing; Liu, Shousheng; Zhao, Zhenzhen; Chen, Lizhen; Jin, Wenwen; Li, Kai; Xin, Yongning; Zhuang, Likun

doi:10.1016/j.omtn.2021.12.017

Cited by 10 publications

(8 citation statements)

References 46 publications

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“…In addition, we found that DRAM1 was prominently expressed in regions at or near the plasma membrane of RAW264.7 macrophages. Plasma membrane expression of DRAM1 has also been observed in hepatocytes and two other members of the DRAM family, DRAM3 and DRAM5, also display plasma membrane expression [24,26,38]. Additionally, we observed the formation of circular DRAM1-positive structures near the plasma membrane that might be the result of endocytosis, for example of cell debris, and therefore derived from the plasma membrane in regions where DRAM1 is highly concentrated.…”

Section: Discussionsupporting

confidence: 68%

“…Additionally, we observed the formation of circular DRAM1-positive structures near the plasma membrane that might be the result of endocytosis, for example of cell debris, and therefore derived from the plasma membrane in regions where DRAM1 is highly concentrated. This expression pattern of DRAM1 is particularly interesting in the light of a recent study reporting a role for DRAM1 in extracellular vesicles formation [38]. Since exosomes are crucial for communication between immune cells, this might point to an unexplored role for DRAM1 in the intercellular propagating of signals during infection.…”

Section: Discussionmentioning

confidence: 75%

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DRAM1 promotes antibacterial autophagy and lysosomal delivery of Mycobacterium marinum in macrophages

Banducci-Karp

Xie

Engels

et al. 2022

Preprint

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Damage-Regulated Autophagy Modulator 1 (DRAM1) is an infection-inducible membrane protein, whose function in the immune response is incompletely understood. Based on previous results in a zebrafish infection model, we have proposed that DRAM1 is a host resistance factor against intracellular mycobacterial infection. To gain insight into the cellular processes underlying DRAM1-mediated host defence, here we studied the interaction of DRAM1 with Mycobacterium marinum in murine RAW264.7 macrophages. We found that shortly after phagocytosis, DRAM1 localised in a punctate pattern to mycobacteria, which gradually progressed to full DRAM1 envelopment of the bacteria. Within the same time frame, DRAM1-positive mycobacteria colocalised with the LC3 marker for autophagosomes and LysoTracker and LAMP1 markers for (endo)lysosomes. Knockdown analysis revealed that DRAM1 is required for recruitment of LC3 and for acidification of mycobacteria-containing vesicles. A reduction in the presence of LAMP1 further suggested reduced fusion of lysosomes with mycobacteria-containing vesicles. Finally, we show that DRAM1 knockdown impairs the ability of macrophages to control mycobacterial infection. Together, these results support that DRAM1 promotes the trafficking of mycobacteria through the degradative (auto)phagolysosomal pathway. Considering its prominent effect on host resistance to intracellular infection, DRAM1 is a promising target for therapeutic modulation of the microbicidal capacity of macrophages.

show abstract

Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 75%

DRAM1 promotes antibacterial autophagy and lysosomal delivery of Mycobacterium marinum in macrophages

Banducci-Karp

Xie

Engels

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Additionally, we observed the formation of circular DRAM1-positive structures near the plasma membrane that might be the result of endocytosis, for example, of cell debris, and that, therefore, derived from the plasma membrane in regions where DRAM1 is highly concentrated. This expression pattern of DRAM1 is particularly interesting in the light of a recent study reporting a role for DRAM1 in extracellular vesicle formation [ 44 ]. Since exosomes are crucial for communication between immune cells, this might point to an unexplored role for DRAM1 in the intercellular propagating of signals during infection.…”

Section: Discussionmentioning

confidence: 88%

“…In addition, we found that DRAM1 was prominently expressed in regions at or near the plasma membrane of RAW264.7 macrophages. Plasma membrane expression of DRAM1 has also been observed in hepatocytes, and two other members of the DRAM family, DRAM3 and DRAM5, also display plasma membrane expression [ 27 , 29 , 44 ]. Additionally, we observed the formation of circular DRAM1-positive structures near the plasma membrane that might be the result of endocytosis, for example, of cell debris, and that, therefore, derived from the plasma membrane in regions where DRAM1 is highly concentrated.…”

Section: Discussionmentioning

confidence: 99%

DRAM1 Promotes Lysosomal Delivery of Mycobacterium marinum in Macrophages

Banducci-Karp

Xie

Engels

et al. 2023

Cells

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Damage-Regulated Autophagy Modulator 1 (DRAM1) is an infection-inducible membrane protein, whose function in the immune response is incompletely understood. Based on previous results in a zebrafish infection model, we have proposed that DRAM1 is a host resistance factor against intracellular mycobacterial infection. To gain insight into the cellular processes underlying DRAM1-mediated host defence, here we studied the interaction of DRAM1 with Mycobacterium marinum in murine RAW264.7 macrophages. We found that, shortly after phagocytosis, DRAM1 localised in a punctate pattern to mycobacteria, which gradually progressed to full DRAM1 envelopment of the bacteria. Within the same time frame, DRAM1-positive mycobacteria colocalised with the LC3 marker for autophagosomes and LysoTracker and LAMP1 markers for (endo)lysosomes. Knockdown analysis revealed that DRAM1 is required for the recruitment of LC3 and for the acidification of mycobacteria-containing vesicles. A reduction in the presence of LAMP1 further suggested reduced fusion of lysosomes with mycobacteria-containing vesicles. Finally, we show that DRAM1 knockdown impairs the ability of macrophages to defend against mycobacterial infection. Together, these results support that DRAM1 promotes the trafficking of mycobacteria through the degradative (auto)phagolysosomal pathway. Considering its prominent effect on host resistance to intracellular infection, DRAM1 is a promising target for therapeutic modulation of the microbicidal capacity of macrophages.

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“…Third, previous works, including ours, have shown that apoptotic or hypoxic stress induce the translocation of PKM2 to the nucleus and cell surface in several cancers 25 , 65 , 66 . Fourth, ethanol or hypoxia increases the expression of PKM2, which is frequently observed in exosomes and ectosomes secreted from the cancer cell surface 67 , 68 . Moreover, our current and previous studies both indicate that hypoxia induces increased PKM2 expression in HCC 25 (Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

Hypoxia-activated selectivity-improved anti-PKM2 antibody combined with prodrug TH-302 for potentiated targeting therapy in hepatocellular carcinoma

Wang,

Qi,

Chen

et al. 2024

Int. J. Biol. Sci.

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Background: Hypoxia induces hepatocellular carcinoma (HCC) malignancies; yet it also offers treatment opportunities, exemplified by developing hypoxia-activated prodrugs (HAPs). Although HAP TH-302 combined with therapeutic antibody (Ab) has synergistic effects, the clinical benefits are limited by the on-target off-tumor toxicity of Ab. Here, we sought to develop a hypoxia-activated anti-M2 splice isoform of pyruvate kinase (PKM2) Ab combined with TH-302 for potentiated targeting therapy. Methods: Codon-optimized and hypoxia-activation strategies were used to develop H103 Ab-azo-PEG5k (HAP103) Ab. Hypoxia-activated HAP103 Ab was characterized, and hypoxia-dependent antitumor and immune activities were evaluated. Selective imaging and targeting therapy with HAP103 Ab were assessed in HCC-xenografted mouse models. Targeting selectivity, systemic toxicity, and synergistic therapeutic efficacy of HAP103 Ab with TH-302 were evaluated. Results: Human full-length H103 Ab was produced in a large-scale bioreactor. Azobenzene (azo)-linked PEG5k conjugation endowed HAP103 Ab with hypoxia-activated targeting features. Conditional HAP103 Ab effectively inhibited HCC cell growth, enhanced apoptosis, and induced antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) functions. Analysis of HCC-xenografted mouse models showed that HAP103 Ab selectively targeted hypoxic HCC tissues and induced potent tumor-inhibitory activity either alone or in combination with TH-302. Besides the synergistic effects, HAP103 Ab had negligible side effects when compared to parent H103 Ab. Conclusion: The hypoxia-activated anti-PKM2 Ab safely confers a strong inhibitory effect on HCC with improved selectivity. This provides a promising strategy to overcome the on-target off-tumor toxicity of Ab therapeutics; and highlights an advanced approach to precisely kill HCC in combination with HAP TH-302.

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DRAM1 increases the secretion of PKM2-enriched EVs from hepatocytes to promote macrophage activation and disease progression in ALD

Cited by 10 publications

References 46 publications

DRAM1 promotes antibacterial autophagy and lysosomal delivery of Mycobacterium marinum in macrophages

DRAM1 promotes antibacterial autophagy and lysosomal delivery of Mycobacterium marinum in macrophages

DRAM1 Promotes Lysosomal Delivery of Mycobacterium marinum in Macrophages

Hypoxia-activated selectivity-improved anti-PKM2 antibody combined with prodrug TH-302 for potentiated targeting therapy in hepatocellular carcinoma

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