A defective lysophosphatidic acid-autophagy axis increases miscarriage risk by restricting decidual macrophage residence

Hui-Li Yang; Zhen-Zhen Lai; Jia-Wei Shi; Wen-Jie Zhou; Jie Mei; Jiang-Feng Ye; Tao Zhang; Jian Wang; Jian-Yuan Zhao; Da-Jin Li; Ming-Qing Li

doi:10.1080/15548627.2022.2039000

A defective lysophosphatidic acid-autophagy axis increases miscarriage risk by restricting decidual macrophage residence

Autophagy. 2022 Oct;18(10):2459-2480. doi: 10.1080/15548627.2022.2039000. Epub 2022 Feb 27.

Authors

Hui-Li Yang^{1

2}, Zhen-Zhen Lai¹, Jia-Wei Shi¹, Wen-Jie Zhou³, Jie Mei⁴, Jiang-Feng Ye⁵, Tao Zhang⁶, Jian Wang¹, Jian-Yuan Zhao^{7

8}, Da-Jin Li¹, Ming-Qing Li^{1

2}

Affiliations

¹ Laboratory for Reproductive Immunology, NHC Key Lab of Reproduction Regulation (Shanghai Institute for Biomedical and Pharmaceutical Technologies), Hospital of Obstetrics and Gynecology, Shanghai Medical School, Fudan University, Shanghai, 200080 People's Republic of China.
² Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Hospital of Obstetrics and Gynecology, Shanghai Medical School, Fudan University, Shanghai, 200080, People's Republic of China.
³ Center of Reproductive Medicine of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, People's Republic of China.
⁴ Reproductive Medicine Center, Department of Obstetrics and Gynecology, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medicine School, Nanjing, 210000, People's Republic of China.
⁵ Division of Obstetrics and Gynecology, KK Women's and Children's Hospital, 229899, Singapore.
⁶ Assisted Reproductive Technology Unit, Department of Obstetrics and Gynecology, Faculty of Medicine, Chinese University of Hong Kong, Hong Kong, People's Republic of China.
⁷ State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
⁸ Institute of Metabolism and Integrative Biology (IMIB), School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.

Abstract

Massive infiltrated and enriched decidual macrophages (dMφ) have been widely regarded as important regulators of maternal-fetal immune tolerance and trophoblast invasion, contributing to normal pregnancy. However, the characteristics of metabolic profile and the underlying mechanism of dMφ residence remain largely unknown. Here, we observe that dMφ display an active glycerophospholipid metabolism. The activation of ENPP2-lysophosphatidic acid (LPA) facilitates the adhesion and retention, and M2 differentiation of dMφ during normal pregnancy. Mechanistically, this process is mediated through activation of the LPA receptors (LPAR1 and PPARG/PPARγ)-DDIT4-macroautophagy/autophagy axis, and further upregulation of multiple adhesion factors (e.g., cadherins and selectins) in a CLDN7 (claudin 7)-dependent manner. Additionally, poor trophoblast invasion and placenta development, and a high ratio of embryo loss are observed in Enpp2^±, lpar1^-/- or PPARG-blocked pregnant mice. Patients with unexplained spontaneous abortion display insufficient autophagy and cell residence of dMφ. In therapeutic studies, supplementation with LPA or the autophagy inducer rapamycin significantly promotes dMφ autophagy and cell residence, and improves embryo resorption in Enpp2^± and spontaneous abortion mouse models, which should be dependent on the activation of DDIT4-autophagy-CLDN7-adhesion molecules axis. This observation reveals that inactivation of ENPP2-LPA metabolism and insufficient autophagy of dMφ result in resident obstacle of dMφ and further increase the risk of spontaneous abortion, and provides potential therapeutic strategies to prevent spontaneous abortion.Abbreviations: ACTB: actin beta; ADGRE1/F4/80: adhesion G protein-coupled receptor E1; Atg5: autophagy related 5; ATG13: autophagy related 13; BECN1: beclin 1; CDH1/E-cadherin: cadherin 1; CDH5/VE-cadherin: cadherin 5; CFSE: carboxyfluorescein succinimidyl ester; CLDN7: claudin 7; CSF1/M-CSF: colony stimulating factor 1; CSF2/GM-CSF: colony stimulating factor 2; Ctrl: control; CXCL10/IP-10: chemokine (C-X-C) ligand 10; DDIT4: DNA damage inducible transcript 4; dMφ: decidual macrophage; DSC: decidual stromal cells; ENPP2/ATX: ectonucleotide pyrophosphatase/phosphodiesterase 2; Enpp2^±: Enpp2 heterozygous knockout mouse; ENPP2i/PF-8380: ENPP2 inhibitor; EPCAM: epithelial cell adhesion molecule; ESC: endometrial stromal cells; FGF2/b-FGF: fibroblast growth factor 2; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GPCPD1: glycerophosphocholine phosphodiesterase 1; HE: heterozygote; HIF1A: hypoxia inducible factor 1 subunit alpha; HNF4A: hepatocyte nuclear factor 4 alpha; HO: homozygote; ICAM2: intercellular adhesion molecule 2; IL: interleukin; ITGAV/CD51: integrin subunit alpha V; ITGAM/CD11b: integrin subunit alpha M; ITGAX/CD11b: integrin subunit alpha X; ITGB3/CD61: integrin subunit beta 3; KLRB1/NK1.1: killer cell lectin like receptor B1; KRT7/cytokeratin 7: keratin 7; LPA: lysophosphatidic acid; LPAR: lysophosphatidic acid receptor; lpar1^-/-: lpar1 homozygous knockout mouse; LPAR1i/AM966: LPAR1 inhibitor; LY6C: lymphocyte antigen 6 complex, locus C1; LYPLA1: lysophospholipase 1; LYPLA2: lysophospholipase 2; Lyz2: lysozyme 2; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MARVELD2: MARVEL domain containing 2; 3-MA: 3-methyladenine; MBOAT2: membrane bound O-acyltransferase domain containing 2; MGLL: monoglyceride lipase; MRC1/CD206: mannose receptor C-type 1; MTOR: mechanistic target of rapamycin kinase; NP: normal pregnancy; PDGF: platelet derived growth factor; PLA1A: phospholipase A1 member A; PLA2G4A: phospholipase A2 group IVA; PLPP1: phospholipid phosphatase 1; pMo: peripheral blood monocytes; p-MTOR: phosphorylated MTOR; PPAR: peroxisome proliferator activated receptor; PPARG/PPARγ: peroxisome proliferator activated receptor gamma; PPARGi/GW9662: PPARG inhibitor; PTPRC/CD45: protein tyrosine phosphatase receptor type, C; Rapa: rapamycin; RHEB: Ras homolog, mTORC1 binding; SA: spontaneous abortion; SELE: selectin E; SELL: selectin L; siCLDN7: CLDN7-silenced; STAT: signal transducer and activator of transcription; SQSTM1: sequestosome 1; TJP1: tight junction protein 1; VCAM1: vascular cell adhesion molecule 1; WT: wild type.

Keywords: Abortion; CLDN7; DDIT4; ENPP2; LPAR1; decidual macrophage; lysophosphatidic acid; trophoblast invasion.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Abortion, Spontaneous* / genetics
Abortion, Spontaneous* / metabolism
Actins / metabolism
Acyltransferases / metabolism
Animals
Autophagy* / genetics
Beclin-1 / metabolism
Cadherins / metabolism
Chemokine CXCL10 / metabolism
Claudins / metabolism
Epithelial Cell Adhesion Molecule / metabolism
Esters / metabolism
Female
Fibroblast Growth Factor 2 / metabolism
Glycerophospholipids / metabolism
Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
Group IV Phospholipases A2 / metabolism
Hepatocyte Nuclear Factor 4 / metabolism
Humans
Hypoxia-Inducible Factor 1 / metabolism
Integrins / metabolism
Keratin-7 / metabolism
Ligands
Lysophospholipase / metabolism
Lysophospholipids / metabolism
MARVEL Domain Containing 2 Protein
Macrophage Colony-Stimulating Factor / metabolism
Macrophages / metabolism
Mechanistic Target of Rapamycin Complex 1 / metabolism
Mice
Mice, Knockout
Microtubule-Associated Proteins / metabolism
Monoacylglycerol Lipases / metabolism
Muramidase / metabolism
PPAR gamma / metabolism
Phospholipases
Phospholipases A1 / metabolism
Phosphoric Diester Hydrolases / genetics
Phosphoric Diester Hydrolases / metabolism
Phosphoric Monoester Hydrolases / metabolism
Platelet-Derived Growth Factor / metabolism
Pregnancy
Pyrophosphatases / metabolism
Receptors, Lysophosphatidic Acid / metabolism
Receptors, NK Cell Lectin-Like / metabolism
Selectins / metabolism
Sequestosome-1 Protein / metabolism
Sirolimus
TOR Serine-Threonine Kinases / metabolism
Thiolester Hydrolases
Vascular Cell Adhesion Molecule-1 / metabolism
Zonula Occludens-1 Protein / metabolism

Substances

Actins
Beclin-1
CLDN7 protein, human
Cadherins
Chemokine CXCL10
Claudins
Epithelial Cell Adhesion Molecule
Esters
Glycerophospholipids
Hepatocyte Nuclear Factor 4
Hypoxia-Inducible Factor 1
Integrins
Keratin-7
Ligands
Lysophospholipids
MARVEL Domain Containing 2 Protein
MARVELD2 protein, human
Microtubule-Associated Proteins
PPAR gamma
Platelet-Derived Growth Factor
Receptors, Lysophosphatidic Acid
Receptors, NK Cell Lectin-Like
Selectins
Sequestosome-1 Protein
Vascular Cell Adhesion Molecule-1
Zonula Occludens-1 Protein
Fibroblast Growth Factor 2
Macrophage Colony-Stimulating Factor
Granulocyte-Macrophage Colony-Stimulating Factor
Acyltransferases
Mechanistic Target of Rapamycin Complex 1
TOR Serine-Threonine Kinases
GPCPD1 protein, human
Phospholipases
Monoacylglycerol Lipases
Phospholipases A1
Group IV Phospholipases A2
Lysophospholipase
Lypla1 protein, mouse
Thiolester Hydrolases
Phosphoric Monoester Hydrolases
Phosphoric Diester Hydrolases
Muramidase
Pyrophosphatases
lysophosphatidic acid
Sirolimus

Grants and funding

This study was supported by the Major Research Program of National Natural Science Foundation of China (NSFC) (No. 92057119, 31970798, 32070915, 81901563 and 82071646); the National Key Research and Development Program of China (2017YFC1001404); the Innovation-oriented Science and Technology Grant from NPFPC Key Laboratory of Reproduction Regulation (CX2017-2); the Program for Zhuoxue of Fudan University (JIF157602) and the Support Project for Original Personalized Research of Fudan University (IDF157014/002>) and the Shanghai Sailing Program (to H.L.Y.).