A new era of macrophage-based cell therapy

Na Yi Rang,Kim Sang Wha,Seok Seung Hyeok 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Macrophages are essential innate immune cells found throughout the body that have protective and pathogenic functions in many diseases. When activated, macrophages can mediate the phagocytosis of dangerous cells or materials and participate in effective tissue regeneration by providing growth factors and anti-inflammatory molecules. Ex vivo-generated macrophages have thus been used in clinical trials as cell-based therapies, and based on their intrinsic characteristics, they outperformed stem cells within specific target diseases. In addition to the old methods of generating naïve or M2 primed macrophages, the recently developed chimeric antigen receptor-macrophages revealed the potential of genetically engineered macrophages for cell therapy. Here, we review the current developmental status of macrophage-based cell therapy. The findings of important clinical and preclinical trials are updated, and patent status is investigated. Additionally, we discuss the limitations and future directions of macrophage-based cell therapy, which will help broaden the potential utility and clinical applications of macrophages.

Bone morphogenetic protein 7 induces mesenchymal-to-epithelial transition in melanoma cells, leading to inhibition of metastasis

Na, Yi-Rang,Seok, Seung-Hyeok,Kim, Dong-Jae,Han, Ju-Hee,Kim, Tae-Hyoun,Jung, Hyun,Lee, Byoung-Hee,Park, Jae-Hak Wiley (Blackwell Publishing) 2009 Cancer Science Vol.100 No.11

<P>Bone morphogenetic protein (BMP) 7 counteracts physiological epithelial-to-mesenchymal transition, a process that is indicative of epithelial plasticity in developmental stages. Because epithelial-to-mesenchymal transition and its reversed process mesenchymal-to-epithelial transition (MET) are also involved in cancer progression, we investigated whether BMP7 plays a role in WM-266-4 melanoma cell growth and metastasis. An MTT assay was conducted in WM-266-4 and HEK293T cell lines to show the cell growth inhibition ability of BMP7 and cisplatin. Semiquantitative RT-PCR was used to determine MET in morphologically changed BMP7-treated melanoma cells. MET-induced cells expressed less a basic helix-loop-helix transcription factor (TWIST) in western blot analysis, and we confirm that BMP receptor (Alk2) siRNA transduction could restore TWIST protein expression via blocking of Smad 1, 5 and 8 signaling. Matrigel invasion and cell migration assays were done to investigate the BMP7-induced metastasis inhibition ability. BMP7 treatment only slightly reduced cell growth rate, but induced apparent MET. BMP7 also reduced the invasion and migration ability. Furthermore, BMP7 reduced the resistance of WM-266-4 cells to cisplatin. Collectively, our findings indicate that the metastatis inhibition ability of BMP7 is involved in MET, and that BMP7 could be used as a potential metastasis inhibitor in human melanoma cells.</P>

Tumor microenvironment and metastasis : Clue from the embryonic zebrafish

Yi-Rang Na 한국실험동물학회 2010 한국실험동물학회 학술발표대회 논문집 Vol.2010 No.8

Zebrafish embryo extracts promote sphere-forming abilities of human melanoma cell line.

Na, Yi-Rang,Seok, Seung-Hyeok,Kim, Dong-Jae,Han, Ju-Hee,Kim, Tae-Hyun,Jung, Hyoun,Park, Jae-Hak Japanese Cancer Association 2009 Cancer Science Vol.100 No.8

<P>Sphere-forming abilities in culture condition are considered a hallmark of cancer stem-like cells, which represents tumor cell invasiveness and stem-like characteristics. We aimed to show that the sphere-forming subpopulation of human malignant melanoma cell line WM-266-4 acts differently to zebrafish embryo extracts compared with their bulk counterpart. Spheres were maintained in neural stem cell culture conditions. The embryos of zebrafish at specific developmental stages were collected and the extracts were purified under 100 kDa. Spheres were treated with embyo extracts and proliferation assay and immunocytochemistry were conducted. Spheroid cells expressed nestin and epidermal growth factor receptor (EGFR) but not melanoma antigen recognized by T-cells (MART)1, indicating their stem-like character. Zebrafish embryo extracts at 50% epiboly stage inhibited melanoma bulk cell proliferation in a dose-dependent manner. However, sphere-forming abilities were significantly enhanced under 1 microg/mL concentration of 50% epiboly stage embryo extract treatment. Our findings implicate that we should consider cell subsets of a different character from the tumor origin that can respond differently to exogenous substances or tumor microenvironments. We suggest that cancer research should consider both minor stem-like subpopulations and the other major bulk tumor cells.</P>

Protective effects of Vitamin E against chlorinated AHR Ligand 3,3`, 4,4`, 5-pentachlorobiphenyl (PCB126) induced toxicity in developmental zebrafish (Danio rerio) embryos

( Yi Rang Na ),( Seung Hyeok Seok ),( Min Won Baek ),( Hui Young Lee ),( Dong Jae Kim ),( Sung Hoon Park ),( Hyun Kyoung Lee ),( Byoung Hee Lee ),( Jae Hak Park ) 한국동물실험대체법학회 2007 한국동물실험대체법학회 학술대회집 Vol.2007 No.1

Pro-oxidant 3,3`,4,4`,5-pentachlorinated biphenyls 126 (PCB126) is a global environmental contaminant that can induce cellular oxidative stress. Disruption from cellular oxidative stress can be blocked by several nutrient antioxidants like vitamin E. We used the fish model, zebrafish (Danio rerio), to determine if vitamin E can protect against toxicity from the aryl hydrocarbon receptor (AHR) ligand PCB126 during zebrafish development, and to show the mechanism by which PCB 126 induces toxic effects. We exposed zebrafish embryos at 8 hours post fertilization (hpf) to PCB126 at a concentration of 100 nM, and compared these fish with a second group that was co-exposed to 100-μM vitamin E and 100-nM PCB126 until 5 days post fertilization (dpf). PCB126 induced pericardial sac edema, yolk sac edema, heart malformation, hemorrhaging, failure of swim bladder inflation, growth retardation and mortality in developing zebrafish larvae. In contrast, 100-μM vitamin E co-exposure did not induce the gross changes seen in the PCB126 treatment group. We found that vitamin E can protect against PCB126 toxicity by inhibiting zebrafish heat shock protein 70 cognate (zfHsc70) down regulation. Also vitamin E inhibited the zebrafish aryl hydrocarbon receptor type-2 (zfAHR2), zebrafish cytochrome P450 1A (zfCYP1A), zebrafish superoxide dismutase-1 (zfSOD1) and upregulated zebrafish superoxide dismutase-2 (zfSOD2) mRNA expression. These data give insights into the use of vitamin E to reduce PCB126-mediated toxicity and into the use of zebrafish embryos for exploring mechanisms underlying the oxidative potential of AHR agonists.

GM-CSF Induces Inflammatory Macrophages by Regulating Glycolysis and Lipid Metabolism

Na, Yi Rang,Gu, Gyo Jung,Jung, Daun,Kim, Young Won,Na, Juri,Woo, Jin Sun,Cho, Joo Youn,Youn, Hyewon,Seok, Seung Hyeok The American Association of Immunologists, Inc. 2016 JOURNAL OF IMMUNOLOGY Vol.197 No.10

<P>GM-CSF induces proinflammatory macrophages, but the underlying mechanisms have not been studied thus far. In this study, we investigated the mechanisms of how GM-CSF induces inflammatory macrophages. First, we observed that GM-CSF increased the extent of LPS-induced acute glycolysis in murine bone marrow-derived macrophages. This directly correlates with an inflammatory phenotype because glycolysis inhibition by 2-deoxyglucose abolished GM-CSF-mediated increase of TNF-alpha,IL-1 beta, IL-6, and IL-12p70 synthesis upon LPS stimulation. Increased glycolytic capacity is due to de novo synthesis of glucose transporter ( GLUT)-1,-3, and-4, as well as c-myc. Mean while, GM-CSF increased 3-hydroxy-3-methyl-glutaryl-CoA reductase, which is the rate-limiting enzyme of the mevalonate pathway. Inhibition of acute glycolysis or 3-hydroxy-3-methyl-glutaryl-CoA reductase abrogated the inflammatory effects of GM-CSF priming in macrophages. Finally, mice with inflamed colons exposed to dextran sodium sulfate containing GLUT-1(high) macrophages led to massive uptake of [F-18]-fluorodeoxyglucose, but GM-CSF neutralization reduced the positron-emission tomography signal in the intestine and also decreased GLUT-1 expression in colonic macrophages. Collectively, our results reveal glycolysis and lipid metabolism created by GM-CSF as the underlying metabolic constructs for the function of inflammatory macrophages.</P>

Proteomic Analysis Reveals Distinct Metabolic Differences Between Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) and Macrophage Colony Stimulating Factor (M-CSF) Grown Macrophages Derived from Murine Bone Marrow Cells

Na, Yi Rang,Hong, Ji Hye,Lee, Min Yong,Jung, Jae Hun,Jung, Daun,Kim, Young Won,Son, Dain,Choi, Murim,Kim, Kwang Pyo,Seok II, Seung Hyeok The American Society for Biochemistry and Molecula 2015 Molecular and Cellular Proteomics Vol.14 No.10

<P>Macrophages are crucial in controlling infectious agents and tissue homeostasis. Macrophages require a wide range of functional capabilities in order to fulfill distinct roles in our body, one being rapid and robust immune responses. To gain insight into macrophage plasticity and the key regulatory protein networks governing their specific functions, we performed quantitative analyses of the proteome and phosphoproteome of murine primary GM-CSF and M-CSF grown bone marrow derived macrophages (GM-BMMs and M-BMMs, respectively) using the latest isobaric tag based tandem mass tag (TMT) labeling and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Strikingly, metabolic processes emerged as a major difference between these macrophages. Specifically, GM-BMMs show significant enrichment of proteins involving glycolysis, the mevalonate pathway, and nitrogen compound biosynthesis. This evidence of enhanced glycolytic capability in GM-BMMs is particularly significant regarding their pro-inflammatory responses, because increased production of cytokines upon LPS stimulation in GM-BMMs depends on their acute glycolytic capacity. In contrast, M-BMMs up-regulate proteins involved in endocytosis, which correlates with a tendency toward homeostatic functions such as scavenging cellular debris. Together, our data describes a proteomic network that underlies the pro-inflammatory actions of GM-BMMs as well as the homeostatic functions of M-BMMs.</P>

GM-CSF Grown Bone Marrow Derived Cells Are Composed of Phenotypically Different Dendritic Cells and Macrophages

Na, Yi Rang,Jung, Daun,Gu, Gyo Jeong,Seok, Seung Hyeok Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.10

Granulocyte-macrophage colony stimulating factor (GM-CSF) has a role in inducing emergency hematopoiesis upon exposure to inflammatory stimuli. Although GM-CSF generated murine bone marrow derived cells have been widely used as macrophages or dendritic cells in research, the exact characteristics of each cell population have not yet been defined. Here we discriminated GM-CSF grown bone marrow derived macrophages (GM-BMMs) from dendritic cells (GM-BMDCs) in several criteria. After C57BL/6J mice bone marrow cell culture for 7 days with GM-CSF supplementation, two main populations were observed in the attached cells based on MHCII and F4/80 marker expressions. GM-BMMs had $MHCII^{low}F4/80^{high}$ as well as $CD11c^+CD11b^{high}CD80^-CD64^+MerTK^+$ phenotypes. In contrast, GM-BMDCs had $MHCII^{high}F4/80^{low}$ and $CD11c^{high}CD8{\alpha}^-CD11b^+CD80^+CD64^-MerTK^{low}$ phenotypes. Interestingly, the GM-BMM population increased but GM-BMDCs decreased in a GM-CSF dose-dependent manner. Functionally, GM-BMMs showed extremely high phagocytic abilities and produced higher IL-10 upon LPS stimulation. GM-BMDCs, however, could not phagocytose as well, but were efficient at producing $TNF{\alpha}$, $IL-1{\beta}$, IL-12p70 and IL-6 as well as inducing T cell proliferation. Finally, whole transcriptome analysis revealed that GM-BMMs and GM-BMDCs are overlap with in vivo resident macrophages and dendritic cells, respectively. Taken together, our study shows the heterogeneicity of GM-CSF derived cell populations, and specifically characterizes GM-CSF derived macrophages compared to dendritic cells.

Metabolic features of macrophages in inflammatory diseases and cancer

Na, Yi Rang,Je, Sungmo,Seok, Seung Hyeok Elsevier 2018 Cancer letters Vol.413 No.-

<P><B>Abstract</B></P> <P>Macrophages are now considered to be important players in various inflammatory diseases as well as tumor progression. Emerging evidence reveals that macrophage metabolic features are deeply associated with their immune functions. Understanding the interaction between cellular metabolism and immune signaling pathways in macrophages can help us to develop appropriate therapeutic approaches for inflammatory diseases. In this review, we briefly summarize key metabolic features of M1 and M2 macrophages as well as signaling interactions between major metabolic molecules with TLRs and NLRs. Current knowledges of cellular metabolism are focused on macrophages in various disease situations including sepsis, atherosclerosis, obesity, tuberculosis and cancer. Novel insights and present targets for regulating macrophage metabolism are also discussed.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Pre-established glycolytic capacity determines inflammatory functions of GM-CSF derived macrophages. </LI> <LI> LPS/IFNγ induces aerobic glycolysis but IL-4/IL-13 potentiates mitochondrial respiration in macrophages. </LI> <LI> AMPK, mTOR and mitochondrial ROS integrate with pattern recognition receptor signaling pathways. </LI> <LI> Key metabolic changes of macrophages were reviewed focused on the inflammatory diseases and cancer. </LI> </UL> </P>

Application of ex-vivo generated macrophages as potential cell therapy

Yi Rang Na 한국실험동물학회 2022 한국실험동물학회 학술발표대회 논문집 Vol.2022 No.7