MFG-E8 Plays an Important Role in Attenuating Cerulein-Induced Acute Pancreatitis in Mice

Bu, Heng‐Fu; Subramanian, Saravanan; Geng, Hua; Wang, Xiao; Liu, Fangyi; Chou, Pauline M.; Du, Chao; Plaen, Isabelle G. De; Tan, Xiaodi

doi:10.3390/cells10040728

Cited by 4 publications

(4 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

Section: Discussionmentioning

confidence: 99%

“…MFG-E8 deficient mice develop inflammation and autoimmunity, including glomerulonephritis, attributed to defects in apoptotic cell removal [ 7 ]. In addition to tag-free rhMFG-E8’s potential to be developed as a biological therapeutic for radiation injury and AKI, it might also be developed for other diseases such as atrial fibrosis and atrial fibrillation [ 46 ], subarachnoid hemorrhage [ 47 , 48 ], osteoarthritis [ 49 ], and acute pancreatitis [ 50 ]. Additionally, MFG-E8 has the potential to become the first example of an entirely new biologic able to promote health and healing by enhancing the removal of dying cells in situations characterized by cytotoxicity, such as infectious diseases, intoxications, envenomations, and in patients undergoing chemotherapy and radiotherapy.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Development of human cell-expressed tag-free rhMFG-E8 as a radiation mitigator and a therapeutic for acute kidney injury

Chaung

Zhang

et al. 2023

Preprint

View full text Add to dashboard Cite

Background Human milk fat globule epidermal growth factor-factor VIII (MFG-E8) functions as a bridging molecule to promote the removal of dying cells by professional phagocytes. E. coli-expressed histidine-tagged recombinant human MFG-E8 (rhMFG-E8) is protective in various disease conditions. However, due to improper recombinant protein glycosylation, misfolding and possible antigenicity, E. coliexpressed histidine-tagged rhMFG-E8 is unsuitable for human therapy. Therefore, we hypothesize that human cell-expressed, tag-free rhMFG-E8 can be developed as a safe and effective novel biologic to treat inflammatory diseases such as radiation injury and acute kidney injury (AKI). Methods We produced a new tag-free rhMFG-E8 protein by cloning the human MFG-E8 full-length coding sequence without any fusion tag into a mammalian vector and expressed it in HEK293-derived cells. The construct includes the leader sequence of cystatin S to maximize secretion of rhMFG-E8 into the culture medium. After purification and confirmation of the protein identity, we first evaluated its biological activity in vitro. We then determined its efficacy in vivo utilizing two experimental rodent models of organ injury: partial body irradiation (PBI) and ischemia/reperfusion-induced AKI. Results HEK293 cell supernatant containing tag-free rhMFG-E8 protein was concentrated, purified, and rhMFG-E8 was verified by SDS-PAGE analysis and mass spectrometry. The biological activity of human cell-expressed tag-free rhMFG-E8 was superior to that of E. coli-expressed His-tagged rhMFG-E8. Toxicity, stability, and pharmacokinetic studies indicate that tag-free rhMFG-E8 is safe, highly stable after lyophilization and long-term storage, and with an adequate half-life for therapeutic applications. In the PBI model, a dose-dependent improvement of the 30-day survival rate was observed after tag-free rhMFG-E8 treatment with a 30-day survival of 89%, which was significantly higher than the 25% survival in the vehicle group. The dose modification factor (DMF) of tag-free rhMFG-E8 was 1.073. Tag-free rhMFG-E8 also attenuated gastrointestinal damage after PBI. In the model of AKI, tag-free rhMFG-E8 treatment significantly attenuated kidney injury and inflammation, and improved the 10-day survival. Conclusion Our new human cell-expressed tag-free rhMFG-E8 can be further developed as a safe and effective therapy to treat victims of severe acute radiation injury and patients with acute kidney injury.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Development of human cell-expressed tag-free rhMFG-E8 as a radiation mitigator and a therapeutic for acute kidney injury

Chaung

Zhang

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…In addition to tag-free rhMFG-E8’s potential to be developed as a biological therapeutic for radiation injury, it might also be developed for other diseases such as atrial fibrosis and atrial fibrillation 51 , subarachnoid hemorrhage 52 , 53 , osteoarthritis 54 , and acute pancreatitis 55 . Additionally, MFG-E8 has the potential to become the first example of an entirely new biologic able to promote health and healing by enhancing the removal of dying cells in situations characterized by cytotoxicity, such as infectious diseases, intoxications, envenomations, and in patients undergoing chemotherapy and radiotherapy.…”

Section: Discussionmentioning

confidence: 99%

Human cell-expressed tag-free rhMFG-E8 as an effective radiation mitigator

Chaung,

Ma,

Jacob

et al. 2023

Sci Rep

View full text Add to dashboard Cite

Human milk fat globule epidermal growth factor-factor VIII (MFG-E8) functions as a bridging molecule to promote the removal of dying cells by professional phagocytes. E. coli-expressed histidine-tagged recombinant human MFG-E8 (rhMFG-E8) is protective in various disease conditions. However, due to improper recombinant protein glycosylation, misfolding and the possibility of antigenicity, E. coli-expressed histidine-tagged rhMFG-E8 is unsuitable for human therapy. Therefore, we hypothesize that human cell-expressed, tag-free rhMFG-E8 will have suitable structural and functional properties to be developed as a safe and effective novel biologic to treat inflammatory diseases including radiation injury. We produced a new tag-free rhMFG-E8 protein by cloning the human MFG-E8 full-length coding sequence without any fusion tag into a mammalian vector and expressed it in HEK293-derived cells. The construct includes the leader sequence of cystatin S to maximize secretion of rhMFG-E8 into the culture medium. After purification and confirmation of the protein identity, we first evaluated its biological activity in vitro. We then determined its efficacy in vivo utilizing an experimental rodent model of radiation injury, i.e., partial body irradiation (PBI). HEK293 cell supernatant containing tag-free rhMFG-E8 protein was concentrated, purified, and rhMFG-E8 was verified by SDS-PAGE with the standard human MFG-E8 loaded as control and, mass spectrometry followed by analysis using MASCOT for peptide mass fingerprint. The biological activity of human cell-expressed tag-free rhMFG-E8 was superior to that of E. coli-expressed His-tagged rhMFG-E8. Toxicity, stability, and pharmacokinetic studies indicate that tag-free rhMFG-E8 is safe, highly stable after lyophilization and long-term storage, and with a terminal elimination half-life in circulation of at least 1.45 h. In the 15 Gy PBI model, a dose-dependent improvement of the 30-day survival rate was observed after tag-free rhMFG-E8 treatment with a 30-day survival of 89%, which was significantly higher than the 25% survival in the vehicle group. The dose modification factor (DMF) of tag-free rhMFG-E8 calculated using probit analysis was 1.058. Tag-free rhMFG-E8 also attenuated gastrointestinal damage after PBI suggesting it as a potential therapeutic candidate for a medical countermeasure for radiation injury. Our new human cell-expressed tag-free rhMFG-E8 has proper structural and functional properties to be further developed as a safe and effective therapy to treat victims of severe acute radiation injury.

show abstract

“…Following this, the samples were sectioned (5 μm) and treated with hematoxylin and eosin (H&E, Solarbio). Histological inspection was implemented through a blinded observer utilizing scoring systems adapted from earlier procedures [8]. The images were obtained utilizing a light microscope (Leica, Wetzlar, Germany) at ×100 magnification.…”

Section: Histological Examinationmentioning

confidence: 99%

Abietic acid ameliorates pancreatic injury in acute pancreatitis by modulating MAPK pathway

Ye,

Wang,

Mao

et al. 2023

Trop. J. Pharm Res

View full text Add to dashboard Cite

Purpose: To examine the effect of abietic acid (AA) in the treatment of acute pancreatitis (AP) in mice.Methods: C57BL/6J mice were randomly assigned to 4 groups: sham, AP, AP + 20 mg/kg AA, and AP + 40 mg/kg AA groups. To induce AP mouse model, the mice received intraperitoneal (IP) injections of cerulein. The extent of pancreatic tissue damage was evaluated by histological examination. Serum ALT, lipase, and amylase levels were determined by commercial kits while TUNEL assay was used to assess the apoptosis of pancreatic cells. The contents of Bax, Caspase-3, Bcl-2, p-ERK/ERK, p-JNK/JNK, and p-P38/P38 in pancreatic tissues were evaluated by western blot while the contents of IL-6, TNF-α, IL-1β, nitrite, MDA, and GSH in the tissues were evaluated by enzyme-linked immunosorbent assay (ELISA) kits.Results: AA relieved pancreatic damage and reduced ALT, lipase, and amylase levels in ceruleintreated AP mouse (p < 0.001). AA repressed apoptosis of pancreatic cells in cerulein-induced AP mouse, and inhibited oxidative stress and inflammatory response in the mice by reducing IL-6, TNF-α, IL-1β, nitrite, and MDA contents; it also enhanced the levels of GSH in the tissues (p < 0.001). In addition, AA inhibited MAPK pathway activity in the mice (p < 0.001).Conclusion: AA ameliorates pancreatic damage, pancreatic cell apoptosis, oxidative stress, and inflammation in cerulein-induced AP mouse by inhibiting MAPK pathway. This study offers a new potential drug for the management of AP and expands the relevant molecular regulatory mechanisms.

show abstract

MFG-E8 Plays an Important Role in Attenuating Cerulein-Induced Acute Pancreatitis in Mice

Cited by 4 publications

References 47 publications

Development of human cell-expressed tag-free rhMFG-E8 as a radiation mitigator and a therapeutic for acute kidney injury

Development of human cell-expressed tag-free rhMFG-E8 as a radiation mitigator and a therapeutic for acute kidney injury

Human cell-expressed tag-free rhMFG-E8 as an effective radiation mitigator

Abietic acid ameliorates pancreatic injury in acute pancreatitis by modulating MAPK pathway

Contact Info

Product

Resources

About