Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

Lim, Seng Han; Joseph, Megan; Winde, Charlotte M. de; Acton, Sophie E.; Simoncelli, Sabrina

doi:10.1098/rsob.220377

Cited by 5 publications

(2 citation statements)

References 33 publications

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“…During embryogenesis, PDPN is essential for the heart and lymphatic system formation [88][89][90][91][92][93] . In adult life, PDPN is physiologically expressed by Lymphatic Endothelial Cells (LECs) and Fibroblastic Reticular Cells (FRCs) in lymph nodes and allows immune cells to adhere to LECs and FRC to facilitate migration and antigen presentation [94][95][96] After MI, PDPN acts as an inflammatory marker and newly acquired PDPN by MSC help them to interact and communicate with recruited and activated immune cells CD11b high which highly express the PDPN receptor named C-type lectin-like receptor 2, CLEC-2 19 . The interaction between PDPN and CLEC-2 activates an inflammatory response in immune cells 97,98 , and we recently showed that blocking the interaction between MSC positive for PDPN (MSC PDPN+ ) and macrophages improved cardiac function and scar composition after infarction 17,97,98 .…”

Section: Podoplanin a Possible Mesenchymal Stromal Cells Inflammatory...mentioning

confidence: 99%

“…Myocardial infarction and heart failure [16][17][18]103,104 Mesenchymal stromal cells Cardiac Fibrosis 56,94,105 Mesenchymal stromal cells and early reactive fibroblasts Thrombi and thrombo-inflammation 97,106 Mesenchymal stromal cells Deep vein thrombosis 19,107 Mesenchymal stromal cells Atherosclerotic lesion 23,[108][109][110] Mesenchymal stromal cells, Smooth muscle cells and macrophages Aneurysm and High arteria shear 85,111 Mesenchymal stromal cells Calcification aortic valves 112 Mesenchymal stromal cells and fibroblast…”

Section: Source Of Fundingmentioning

confidence: 99%

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Podoplanin Positive Cell-derived Extracellular Vesicles Contribute to Cardiac Amyloidosis After Myocardial Infarction

Cimini,

Hansmann,

Gonzalez

et al. 2024

Preprint

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BackgroundAmyloidosis is a major long-term complication of chronic disease; however, whether it represents one of the complications of post-myocardial infarction (MI) is yet to be fully understood.MethodsUsing wild-type and knocked-out MI mouse models and characterizing in vitro the exosomal communication between bone marrow-derived macrophages and activated mesenchymal stromal cells (MSC) isolated after MI, we investigated the mechanism behind Serum Amyloid A 3 (SAA3) protein overproduction in injured hearts.ResultsHere, we show that amyloidosis occurs after MI and that amyloid fibers are composed of macrophage-derived SAA3 monomers. SAA3 overproduction in macrophages is triggered by exosomal communication from a subset of activated MSC, which, in response to MI, acquire the expression of a platelet aggregation-inducing type I transmembrane glycoprotein named Podoplanin (PDPN). Cardiac MSCPDPN+communicate with and activate macrophages through their extracellular vesicles or exosomes. Specifically, MSCPDPN+derived exosomes (MSCPDPN+Exosomes) are enriched in SAA3 and exosomal SAA3 protein engages with Toll-like receptor 2 (TRL2) on macrophages, triggering an overproduction and impaired clearance of SAA3 proteins, resulting in aggregation of SAA3 monomers as rigid amyloid deposits in the extracellular space. The onset of amyloid fibers deposition alongside extra-cellular-matrix (ECM) proteins in the ischemic heart exacerbates the rigidity and stiffness of the scar, hindering the contractility of viable myocardium and overall impairing organ function. Using SAA3 and TLR2 deficient mouse models, we show that SAA3 delivered by MSCPDPN+exosomes promotes post-MI amyloidosis. Inhibition of SAA3 aggregation via administration of a retro-inverso D-peptide, specifically designed to bind SAA3 monomers, prevents the deposition of SAA3 amyloid fibrils, positively modulates the scar formation, and improves heart function post-MI.ConclusionOverall, our findings provide mechanistic insights into post-MI amyloidosis and suggest that SAA3 may be an attractive target for effective scar reversal after ischemic injury and a potential target in multiple diseases characterized by a similar pattern of inflammation and amyloid deposition.NOVELTY AND SIGNIFICANCEWhat is known?Accumulation of rigid amyloid structures in the left ventricular wall impairs ventricle contractility.After myocardial infarction cardiac Mesenchymal Stromal Cells (MSC) acquire Podoplanin (PDPN) to better interact with immune cells.Amyloid structures can accumulate in the heart after chronic inflammatory conditions.What information does this article contribute?Whether accumulation of cumbersome amyloid structures in the ischemic scar impairs left ventricle contractility, and scar reversal after myocardial infarction (MI) has never been investigated.The pathophysiological relevance of PDPN acquirement by MSC and the functional role of their secreted exosomes in the context of post-MI cardiac remodeling has not been investigated.Amyloid structures are present in the scar after ischemia and are composed of macrophage-derived Serum Amyloid A (SAA) 3 monomers, although mechanisms of SAA3 overproduction is not established.SUMMARY OF NOVELTY AND SIGNIFICANCEHere, we report that amyloidosis, a secondary phenomenon of an already preexisting and prolonged chronic inflammatory condition, occurs after MI and that amyloid structures are composed of macrophage-derived SAA3 monomers. Frequently studied cardiac amyloidosis are caused by aggregation of immunoglobulin light chains, transthyretin, fibrinogen, and apolipoprotein in a healthy heart as a consequence of systemic chronic inflammation leading to congestive heart failure with various types of arrhythmias and tissue stiffness. Although chronic MI is considered a systemic inflammatory condition, studies regarding the possible accumulation of amyloidogenic proteins after MI and the mechanisms involved in that process are yet to be reported. Here, we show that SAA3 overproduction in macrophages is triggered in a Toll-like Receptor 2 (TLR2)-p38MAP Kinase-dependent manner by exosomal communication from a subset of activated MSC, which, in response to MI, express a platelet aggregation-inducing type I transmembrane glycoprotein named Podoplanin. We provide the full mechanism of this phenomenon in murine models and confirm SAA3 amyloidosis in failing human heart samples. Moreover, we developed a retro-inverso D-peptide therapeutic approach, “DRI-R5S,” specifically designed to bind SAA3 monomers and prevent post-MI aggregation and deposition of SAA3 amyloid fibrils without interfering with the innate immune response.

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Section: Podoplanin a Possible Mesenchymal Stromal Cells Inflammatory...mentioning

confidence: 99%

Section: Source Of Fundingmentioning

confidence: 99%

Podoplanin Positive Cell-derived Extracellular Vesicles Contribute to Cardiac Amyloidosis After Myocardial Infarction

Cimini,

Hansmann,

Gonzalez

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

Podoplanin and its multifaceted roles in mammalian developmental program

Cheok,

Tan,

Chan

et al. 2024

Cells & Development

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Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

et al. 2023

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Upon initial immune challenge, dendritic cells (DCs) migrate to lymph nodes and interact with fibroblastic reticular cells (FRCs) via C-type lectin-like receptor 2 (CLEC-2). CLEC-2 binds to the membrane glycoprotein podoplanin (PDPN) on FRCs, inhibiting actomyosin contractility through the FRC network and permitting lymph node expansion. The hyaluronic acid receptor CD44 is known to be required for FRCs to respond to DCs but the mechanism of action is not fully elucidated. Here, we use DNA-PAINT, a quantitative single molecule super-resolution technique, to visualize and quantify how PDPN clustering is regulated in the plasma membrane of FRCs. Our results indicate that CLEC-2 interaction leads to the formation of large PDPN clusters (i.e. more than 12 proteins per cluster) in a CD44-dependent manner. These results suggest that CD44 expression is required to stabilize large pools of PDPN at the membrane of FRCs upon CLEC-2 interaction, revealing the molecular mechanism through which CD44 facilitates cellular crosstalk between FRCs and DCs.

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Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

Cited by 5 publications

References 33 publications

Podoplanin Positive Cell-derived Extracellular Vesicles Contribute to Cardiac Amyloidosis After Myocardial Infarction

Podoplanin Positive Cell-derived Extracellular Vesicles Contribute to Cardiac Amyloidosis After Myocardial Infarction

Podoplanin and its multifaceted roles in mammalian developmental program

Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

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