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Martinez-Martinez, P.

Publications and source records attributed to Martinez-Martinez, P..

2 recordsLinked to original sources

Complement inhibition by C3-siRNA treatment prevents AChR loss and reduces complement activation in the rat Passive Transfer Myasthenia Gravis (PTMG)

Myasthenia gravis (MG), a well understood antibody-mediated autoimmune disease, is characterized by skeletal muscle weakness and fatigue. The autoantibodies are directed against neuromuscular junction (NMJ) proteins such as the acetylcholine receptor (AChR), essential for neuromuscular signal transmission. Anti-AChR antibodies are mainly of the immunoglobulin (Ig) types IgG1 and IgG3, which have a high capacity to activate the classical complement pathway. In AChR-MG, complement activation and the subsequent formation of the membrane attack complex (MAC) at the NMJ is one of the key effector mechanisms, causing substantial damage to the entire postsynaptic membrane and resulting in the typical clinical manifestations of muscle weakness and fatigue observed in both patients and animal models. Since classical treatment strategies, which focus on general immunosuppression and enhancing the effect of ACh binding to the AChR, do not lead to full remission of symptoms in many cases, new treatment approaches are focusing more specifically on targeting effector mechanisms, such as complement activation. The first complement inhibitor approved for MG was eculizumab, a monoclonal antibody targeting C5. However, clinical trials showed that up to 28% of patients treated with eculizumab did not experience an improvement in myasthenic symptoms, emphasizing the urgent need for alternative therapies. Intervening earlier in the complement cascade might be more beneficial, as it could prevent the release of anaphylatoxins, such as C3a, that contribute to further inflammation at the NMJ. In this study, the passive transfer myasthenia gravis (PTMG) rat model was used to investigate whether silencing hepatic C3 expression with a C3-targeted, small interfering (si)RNA, could ameliorate disease symptoms in the acute phase of MG. Female Lewis rats were injected subcutaneously with different dosages of C3-siRNA prior to PTMG induction with mAb35. C3-siRNA, administered weekly at a dose of 30 mg/kg, significantly prevented weight and AChR loss, and accordingly improved muscle function, as measured by muscle strength tests and electromyography. Consistently, the treatment reduced MAC deposition at the NMJ. Overall, these results provide insight into the efficacy of complement inhibitors in the acute phase of MG and suggest potential strategies for advanced treatment options in AChR-MG crisis.

immunology↗

Isolation and characterization of rare circulating autoantibody-producing cells from patients with muscle-specific kinase myasthenia gravis

Myasthenia gravis (MG) is a chronic autoimmune disorder characterized by muscle weakness and caused by autoantibodies that bind to functional membrane proteins at the neuromuscular junction. Most patients have autoantibodies to the acetylcholine receptor (AChR), but a subset of patients instead have autoantibodies to muscle specific tyrosine kinase (MuSK). MuSK is an essential component of the Agrin/Lipoprotein receptor-related protein 4(LRP4)/MuSK/downstream of tyrosine kinase 7 (DOK7) pathway that is responsible for synaptic differentiation, including clustering of AChRs at the neuromuscular junction, both during development and in adult muscle. Nerve-released Agrin binds to LRP4 which then binds to MuSK, stimulating autophosphorylation and recruitment of DOK7 to complete the membrane component of the pathway. Serum-derived IgG4 subclass MuSK autoantibodies prevent the binding of LRP4 to MuSK, subsequently impairing autophosphorylation, resulting in the loss of Agrin-induced AChR clustering in the mouse myotube-forming C2C12 line. Although this autoimmune mechanism appears well understood, MuSK autoantibodies from patients are polyclonal. Most are IgG4 but IgG1, 2 and 3 are also present and can achieve similar results on AChR clustering. In addition, most bind the first Ig-like domain in MuSK, however some patients harbor serum autoantibodies that recognize other domains in MuSK. We sought to establish individual MuSK IgG clones so that the disease mechanisms could be better understood. We isolated MuSK autoantibody-expressing B cells from MuSK MG patients with a fluorescent-tagged MuSK antigen multimer and generated a panel of human monoclonal autoantibodies (mAbs) from these cells. We produced 77 mAbs from single B cells collected from six MuSK MG patients. Here we focused on three highly specific mAbs that bound quantitatively to MuSK in solution, to MuSK-expressing HEK cells and at mouse neuromuscular junctions where they co-localized with AChRs. These three IgG isotype mAbs (two IgG4 and one IgG3 subclass) recognized the Ig-like domain 2 of MuSK. The three MuSK mAbs inhibited Agrin induced-AChR clustering in C2C12 myotubes, but intriguingly, they enhanced rather than inhibited MuSK phosphorylation. This approach for ex vivo isolation of MuSK MG autoantibody-producing cells and production of human recombinant mAbs has identified distinct autoantibody specificities and likely divergent effector mechanisms. Collectively, these findings will enable a better understanding of MuSK autoantibody-mediated pathology.

immunology↗