LRRK2 mediates microglial neurotoxicity via NFATc2 in rodent models of synucleinopathies

Probably not a substantial claim: may not pass S3 (load-bearing).

Microglia activation is associated with pro-inflammatory cytokine production.

“Although microglia activation is associated with pro-inflammatory cytokine production, activation may also lead to the release of neurotrophic molecules such as brain-derived neurotrophic factor (64).”

• cites [64] Trang T, Beggs S, Salter MW, Brain-derived neurotrophic factor from microglia: a molecular substrate for neuropathic pain. Neuron Glia Biol 7, 99– 108 (2011). 22613083 PMC3748035 sourceCould not verify

  The Introduction describes activated microglia releasing BDNF, but does not say that microglial activation is associated with pro-inflammatory cytokine production. Searches for “pro-inflammatory cytokines” and “cytokine production” found no passage addressing that claim. (LLM, conf=0.72)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

In the canonical NFAT pathway, Ca2+/Calcineurin mediates nuclear translocation of NFAT through the dephosphorylation of multiple serine residues in the N-terminus region, including the nuclear localization signal.

“In the canonical NFAT pathway, Ca2+/Calcineurin mediates nuclear translocation of NFAT through the dephosphorylation of multiple serine residues in the N-terminus region, including the nuclear localization signal (34, 59, 60).”

• cites [34] Crabtree GR, Olson EN, NFAT signaling: choreographing the social lives of cells. Cell 109 Suppl, S67– 79 (2002). 11983154 sourceNot checked — no full text of the source
• cites [59] Feske S, Calcium signalling in lymphocyte activation and disease. Nat Rev Immunol 7, 690– 702 (2007). 17703229 sourceNot checked — no full text of the source
• cites [60] Hogan PG, Chen L, Nardone J, Rao A, Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev 17, 2205– 2232 (2003). 12975316 sourcePartly supported

  The ‘Ca²⁺ and calcineurin’ section says calcineurin dephosphorylates serines in NFAT’s N-terminal regulatory domain, triggering nuclear accumulation. The ‘Other considerations’ section links phosphorylation to exposure of the nuclear localization signal, but the copy does not say the signal itself is dephosphorylated; the Figure 2 caption identifies an SRR-2 phosphate that calcineurin does not remove. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

TLR2 neutralization may also impact neurotrophin production in this mouse model of synucleinopathy.

“We have previously demonstrated that although TLR2 neutralization decreases the expression of pro-inflammatory cytokines it may also impact neurotrophin production in this mouse model of synucleinopathy (7).”

• cites [7] Kim C, Ho DH, Suk JE, You S, Michael S, Kang J, Joong Lee S, Masliah E, Hwang D, Lee HJ, Lee SJ, Neuron-released oligomeric alpha-synuclein is an endogenous agonist of TLR2 for paracrine activation of microglia. Nat Commun 4, 1562 (2013). 23463005 PMC4089961 sourcePartly supported

  The Results section on α-synuclein-induced microglial activation reports that a TLR2-blocking antibody eliminated induced cytokine gene expression in isolated microglia; the in vivo Results report reduced microglial activation in Tlr2-deficient mice. Our copy does not report an effect of TLR2 neutralization on neurotrophin production in a mouse model of synucleinopathy (pages not provided). (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

A more recent study demonstrated an induction of NFAT activity in HEK293T cells expressing LRRK2 or LRRK2 G2019S.

“A more recent study, on the other hand, demonstrated an induction of NFAT activity in HEK293T cells expressing LRRK2 or LRRK2 G2019S (38).”

• cites [38] Takagawa T, Kitani A, Fuss I, Levine B, Brant SR, Peter I, Tajima M, Nakamura S, Strober W, An increase in LRRK2 suppresses autophagy and enhances Dectin-1-induced immunity in a mouse model of colitis. Sci Transl Med 10, (2018). PMC6636639 29875204 sourcePartly supported

  The Results section, “LRRK2 regulates NFAT transcriptional activity in mouse BMDCs” (page unavailable), reports increased NFAT reporter activity in HEK293T cells cotransfected with an LRRK2-expressing plasmid. It does not report the same result for LRRK2 G2019S; a search for G2019S found no relevant passage in our copy. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

NFAT has been considered to be an immune-related protein and expressed in T cells and macrophages.

“NFAT has been considered to be an immune-related protein and expressed in T cells and macrophages (66).”

• cites [66] Vaeth M, Feske S, NFAT control of immune function: New Frontiers for an Abiding Trooper. F1000Res 7, 260 (2018). 29568499 PMC5840618 sourcePartly supported

  The Introduction identifies NFAT as a transcription-factor family first found in activated T cells, and the section on peripheral immune tolerance describes its roles in immune function. The passages in our copy do not establish the claim that NFAT is expressed in macrophages (searched: macrophage, macrophages NFAT expression). (LLM, conf=0.85)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

Microglia activation may also lead to the release of neurotrophic molecules such as brain-derived neurotrophic factor.

“Although microglia activation is associated with pro-inflammatory cytokine production, activation may also lead to the release of neurotrophic molecules such as brain-derived neurotrophic factor (64).”

• cites [64] Trang T, Beggs S, Salter MW, Brain-derived neurotrophic factor from microglia: a molecular substrate for neuropathic pain. Neuron Glia Biol 7, 99– 108 (2011). 22613083 PMC3748035 sourceSupported

May combine two claims.

Extracellular α-synuclein aggregates can interact with microglia via TLR2 to activate the LRRK2-NFATc2 pathway and produce a pro-inflammatory phenotype, and can also interact with other receptors to initiate LRRK2-dependent aggregate phagocytosis and clearance via lysosomal pathways.

“Extracellular α-synuclein aggregates may interact with microglia in at least two different ways: on one hand, they may stimulate TLR2 to activate the LRRK2-NFATc2 pathway and produce a pro-inflammatory phenotype, as shown in the present study; on the other, they can also interact with other receptors to initiate LRRK2-dependent aggregate phagocytosis (48) and clearance via lysosomal pathways (49,”

• cites [48] Marker DF, Puccini JM, Mockus TE, Barbieri J, Lu SM, Gelbard HA, LRRK2 kinase inhibition prevents pathological microglial phagocytosis in response to HIV-1 Tat protein. J Neuroinflammation 9, 261 (2012). 23190742 PMC3538654 sourcePartly supported

  Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured
• cites [49] Lee HJ, Khoshaghideh F, Patel S, Lee SJ, Clearance of alpha-synuclein oligomeric intermediates via the lysosomal degradation pathway. J Neurosci 24, 1888– 1896 (2004). 14985429 PMC6730405 sourceNot checked — no full text of the source
• cites [50] Bae EJ, Lee HJ, Rockenstein E, Ho DH, Park EB, Yang NY, Desplats P, Masliah E, Lee SJ, Antibody-aided clearance of extracellular alpha-synuclein prevents cell-to-cell aggregate transmission. J Neurosci 32, 13454– 13469 (2012). 23015436 PMC3752153 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Different neuronal cell types express different NFAT isoforms.

“However, recent studies have shown that neurons also express NFAT and that different neuronal cell types express different NFAT isoforms (67–70).”

• cites [67] Luo J, Sun L, Lin X, Liu G, Yu J, Parisiadou L, Xie C, Ding J, Cai H, A calcineurin- and NFAT-dependent pathway is involved in alpha-synuclein-induced degeneration of midbrain dopaminergic neurons. Hum Mol Genet 23, 6567– 6574 (2014). 25051958 PMC4240205 sourceSupported
• cites [68] Nguyen T, Di Giovanni S, NFAT signaling in neural development and axon growth. Int J Dev Neurosci 26, 141– 145 (2008). 18093786 PMC2267928 sourcePartly supported

  The Introduction describes NFATc4 activity in hippocampal neurons, and the References list a study of NFATc1 in sympathetic ganglion neurons. These passages support examples of different isoforms in different neuronal populations, but the review does not establish a general pattern of cell-type-specific isoform expression. No page numbers were provided in our copy. (LLM, conf=0.76)

  Experimental — accuracy not yet measured
• cites [69] Graef IA, Wang F, Charron F, Chen L, Neilson J, Tessier-Lavigne M, Crabtree GR, Neurotrophins and netrins require calcineurin/NFAT signaling to stimulate outgrowth of embryonic axons. Cell 113, 657– 670 (2003). 12787506 sourceNot checked — no full text of the source
• cites [70] Vihma H, Luhakooder M, Pruunsild P, Timmusk T, Regulation of different human NFAT isoforms by neuronal activity. J Neurochem 137, 394– 408 (2016). 26851544 sourceNot checked — no full text of the source

May combine two claims.

In aged microglia, αSCM exposure induces NFATc2 in wild type but not Lrrk2−/− microglia, while increasing phosphorylation of NFκB and p38 MAPK in both genotypes.

“similar to postnatal microglia, exposure to αSCM induced NFATc2 in aged wild type but not in aged Lrrk2−/− microglia (fig. S8B).”

• Also stated in: § NFATc2 mediates LRRK2 signaling in microglia exposed to neuron-released α-synuclein · plain

  “In contrast, αSCM treatment increased the phosphorylation of NFκB and p38 MAPK in both aged wild type and Lrrk2−/− microglia (fig. S8B).”

May combine two claims.

NFκB phosphorylation is increased in αSCM-treated wild type microglia but is not affected by Lrrk2 gene depletion, suggesting LRRK2 affects NFATc2 specifically.

“Whereas NFκB phosphorylation was also increased in αSCM-treated wild type microglia, it was not affected by Lrrk2 gene depletion, suggesting that LRRK2 affects NFATc2 specifically (Fig. 3D).”

May combine two claims.

TLR2 blockade with T2.5 inhibits NFATc2 induction by extracellular α-synuclein in wild type microglia, whereas calcineurin inhibitor Cyclosporin A has no effect.

“pretreatment with T2.5 to block TLR2 activity was sufficient to inhibit NFATc2 induction by extracellular α-synuclein in wild type microglia (fig. S6A), whereas an inhibitor of calcineurin, Cyclosporin A, had no effect (fig. S7).”

May combine two claims.

αSCM exposure causes a time-dependent increase in nuclear translocation of NFATc2 in mouse microglia, which is inhibited by LRRK2 kinase inhibitor HG-10-102-01 or genetic depletion of Lrrk2.

“We observed a time-dependent increase in the nuclear translocation of NFATc2 following αSCM exposure, which was significantly (P < 0.05) inhibited by either treatment with LRRK2 kinase inhibitor HG-10-102-01 (100 nM) (Fig. 4B) or genetic depletion of Lrrk2 (Fig. 4C).”

May combine two claims.

Microglial conditioned medium from αSCM-treated wild type microglia (Lrrk2+/+-αS-MgCM) decreases DNA content and viable primary neurons, whereas medium from Lrrk2−/− microglia does not cause neuronal loss.

“Treatment with Lrrk2+/+-αS-MgCM significantly (P < 0.001) decreased both DNA content and the number of viable primary neurons, while neuronal cytotoxicity was not observed in control and Lrrk2+/+-LZ-MgCM-treated cells (Fig. 2, L and M).”

• Also stated in: § LRRK2 modulates neuron-released α-synuclein-mediated microglial neurotoxicity · plain

  “Neuronal loss was not observed in either Lrrk2−/−-αS-MgCM or Lrrk2−/−-LZ-MgCM treatment group (Fig. 2, L and M).”

May combine two claims.

Transcriptome analysis of primary rat microglia exposed to αSCM identifies 213 upregulated genes, of which 43 involved in Toll-like receptor signaling and other immune signaling pathways were selected to reconstruct a signaling network.

“Among the 213 genes upregulated after exposure to αSCM, we selected 43 that were involved in Toll-like receptor signaling and other immune signaling pathways (see details in Supplementary Materials) and reconstructed a signaling network describing their interactions utilizing functional gene enrichment analysis and information on protein-protein interactions (Fig. 3A).”

May combine two claims.

In wild type mouse microglia, total Nfatc2 expression and NFATc2 phosphorylation at S54 increase acutely with αSCM exposure.

“In wild type microglia, total Nfatc2 expression acutely increased with exposure time to αSCM (Fig. 3C).”

• Also stated in: § NFATc2 mediates LRRK2 signaling in microglia exposed to neuron-released α-synuclein · hedged

  “NFATc2 phosphorylation at S54 was also increased at early time points following αSCM exposure, indicating enhanced transcriptional activity (Fig. 3C).”

May combine two claims.

In G2019S KI microglia, cytoplasmic NFATc2 is lower and nuclear NFATc2 is higher than in wild type or Lrrk2−/− microglia.

“Cytoplasmic NFATc2 was significantly (P < 0.001) lower in G2019S KI than in wild type or in Lrrk2−/− microglia, while nuclear NFATc2 was correspondingly higher (Fig. 4D).”

May combine two claims.

Recombinant LRRK2 phosphorylates NFATc2 in vitro, with increased phosphorylation by G2019S LRRK2 and no phosphorylation by kinase-dead D1994A LRRK2.

“Phosphorylation of NFATc2 was rapidly induced by LRRK2 and the amount of phosphorylation was increased with the G2019S mutant (Fig. 5A).”

• Also stated in: § NFATc2 is a kinase substrate for LRRK2 · plain

  “In contrast, NFATc2 was not phosphorylated by the LRRK2 kinase dead mutant D1994A (Fig. 5A).”

May combine two claims.

Recombinant TNFα and IL-6 at concentrations similar to those in Lrrk2+/+-αS-MgCM induce neurotoxicity, and together produce a synergistic effect reproducing the neurotoxicity of Lrrk2+/+-αS-MgCM.

“Whereas treatment with a single cytokine induced neurotoxicity, treatment of both TNFα and IL-6 produced a synergistic effect to reproduce the neurotoxicity observed in Fig. 2, L and M (fig. S5, D and E).”

May combine two claims.

LC-MS/MS identifies five NFATc2 phosphopeptides with higher XIC values when incubated with G2019S-LRRK2 than wildtype LRRK2, and lower abundance with D1994A LRRK2.

“Among the 10 phosphorylated residues that were detected, five corresponding phosphopeptides had XIC values higher when NFATc2 was incubated with G2019S-LRRK2 than when incubated with wildtype LRRK2. The same sites were comparatively less abundant in NFATc2 incubated with LRRK2 D1994A (Fig. 5, B and C).”

May combine two claims.

Structural analysis places NFATc2 T483 in the REL-homology region, T483 and T733 close to the DNA binding surface, and T862, T870, and T893 in the flexible C-terminal region associated with NFATc2 translocation.

“Analysis of publicly available crystal structures of NFATc2 revealed that T483 is located in the REL-homology region (aa 408–677), T483 and T733 are located close to the DNA binding surface (PDB: 1P7H and 1OWR) (40), and the remaining three threonine (T862, T870, and T893) are present in the flexible C-terminal region (aa 678–925) associated with NFATc2 translocation (Fig. 6A) (41).”

• cites [40] Giffin MJ, Stroud JC, Bates DL, von Koenig KD, Hardin J, Chen L, Structure of NFAT1 bound as a dimer to the HIV-1 LTR kappa B element. Nat Struct Biol 10, 800– 806 (2003). 12949493 sourceNot checked — no full text of the source
• cites [41] Stroud JC, Chen L, Structure of NFAT bound to DNA as a monomer. J Mol Biol 334, 1009– 1022 (2003). 14643663 sourceNot checked — no full text of the source

May combine two claims.

Genetic deletion of Lrrk2 does not affect αSCM-induced Il-1β or Ccl2 expression, while enhancing Il-10, inos, Cxcl1, and Ccl5 expression.

“Il-1β and Ccl2 were unaffected by Lrrk2 gene deletion, whereas Il-10, inos, Cxcl1, and Ccl5 were enhanced (Fig. 2, C to H).”

May combine two claims.

In control cases NFATc2 immunoreactivity is abundant in the cytoplasm, whereas in PD/DLB patients NFATc2 is concentrated in the nucleus.

“Whereas control cases displayed abundant NFATc2 immunoreactivity in the cell cytoplasm, in patients with PD/DLB, NFATc2 was concentrated in the nucleus (Fig. 7, B and E).”

May combine two claims.

Pharmacological inhibition of LRRK2 kinase activity attenuates αSCM-induced expression of Tnfα and Il-6, but does not affect Il-1β expression.

“pharmacological inhibition of LRRK2 kinase activity attenuated αSCM-induced expression of Tnfα and Il-6, whereas Il-1β expression was unaffected (fig. S4, A to C).”

May combine two claims.

α-syn transgenic mice show marked induction of Nfatc2 that is rescued by LRRK2 kinase inhibition.

“α-syn transgenic mice showed a marked induction of Nfatc2 and this could be rescued by LRRK2 kinase inhibition (Fig. 8Q).”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

May combine two claims.

Administration of LRRK2 kinase inhibitor at 9 months of age completely restores motor behavioral and motor learning deficits in α-syn transgenic mice, but does not rescue motor neuromuscular impairment.

“Administration of LRRK2 kinase inhibitor at 9 months of age completely restored motor behavioral and motor learning deficits in α-syn transgenic mice (Fig. 8, K and L), whereas motor neuromuscular impairment was not rescued (Fig. 8M).”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

May combine two claims.

LRRK2 inhibition fails to improve non-motor symptoms and worsens them in the α-syn transgenic mouse model.

“However, LRRK2 inhibition not only failed to improve non-motor symptoms, but also worsened them.”

May combine two claims.

Nuclear NFATc2 is increased in neurons, astrocytes, and microglia of α-syn transgenic mice and is normalized by LRRK2 kinase inhibition.

“Double-immunostaining analysis also revealed an increase of nuclear NFATc2 in the neurons, astrocytes, and microglia of α-syn transgenic mice that was normalized by LRRK2 kinase inhibition (fig. S11, A to C).”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

May combine two claims.

Expression of Il-10, Cxcl1, Ccl2, and Ccl4 is not modulated in α-syn transgenic mice and is not affected by LRRK2 inhibitor administration.

“The expression of Il-10, Cxcl1, Ccl2, and Ccl4 were not modulated in α-syn transgenic and were not affected by LRRK2 inhibitor administration (Fig. 8, E to I).”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

May combine two claims.

Egr2 expression is increased in α-syn transgenic mice and is significantly attenuated by LRRK2 kinase inhibitor injection.

“we observed an increase of early growth response 2 (Egr2) expression, a well-established NFATc2 target gene, in α-syn-tg mice, which was also significantly (P < 0.01) attenuated by injection with LRRK2 kinase inhibitor (Fig. 8J).”

• AssumptionRests on an assumption: Egr2 is an NFATc2 target gene.
• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

May combine two claims.

α-syn transgenic brains show increased expression of Tnfα, Il-6, and Il-1β, and LRRK2 kinase inhibitor reduces Tnfα and Il-6 but not Il-1β expression.

“α-syn transgenic brains showed increased expression of Tnfα, Il-6, and Il-1β, and administration of the LRRK2 kinase inhibitor reduced the expression of Tnfα and Il-6 but not Il-1β (Fig. 8, B to D).”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

May combine two claims.

PD/DLB brains showed greater overall expression and nuclear translocation of NFATc2.

“Interestingly, PD/DLB brains showed greater overall expression and nuclear translocation of NFATc2.”

May combine two claims.

A LRRK2 and NFATc2 signaling pathway activated by extracellular α-synuclein exists in acutely isolated microglia from aged mice and is independent of NFκB.

“As such, these results support the existence of a LRRK2 and NFATc2 signaling pathway activated by extracellular α-synuclein and independent of NFκB in acutely isolated microglia from aged mice.”

May combine two claims.

LRRK2 directly interacts with NFATc2 in the cytosol of microglia and indirectly modulates NFATc2 nuclear translocation.

“These experiments collectively suggest that LRRK2 directly interacts with NFATc2 in the cytosol of microglia to then, indirectly, modulate NFATc2 nuclear translocation.”

May combine two claims.

The interaction between LRRK2 and NFATc2 may be critical for extracellular α-synuclein-mediated microglial neuroinflammation, which in turn is essential for neurotoxicity in synucleinopathies.

“The interaction between LRRK2 and NFATc2 may be critical for extracellular α-synuclein-mediated microglial neuroinflammation, which in turn is essential for neurotoxicity in synucleinopathies.”

May combine two claims.

Administration of LRRK2 kinase inhibitor ameliorates neuroinflammation, toxicity, and motor behavior deficits in a mouse model of synucleinopathy, suggesting that targeting LRRK2 and NFATc2 interactions might be of therapeutic interest for synucleinopathies.

“Moreover, administration of LRRK2 kinase inhibitor ameliorated neuroinflammation, toxicity, and motor behavior deficits in this mouse model of synucleinopathy, suggesting that targeting LRRK2 and NFATc2 interactions might be of therapeutic interest for synucleinopathies (fig. S12).”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

Reducing microglial clearance activity might allow α-synuclein to accumulate in the extracellular space.

“Nevertheless, we certainly cannot ignore the concern that reducing microglial clearance activity might allow α-synuclein to accumulate in the extracellular space.”

May combine two claims.

Blocking NFATc2 phosphorylation by LRRK2 inhibition may prevent toxic inflammation but also impede α-synuclein clearance.

“Thus, blocking NFATc2 phosphorylation by LRRK2 inhibition may not only prevent toxic inflammation but also impede α-synuclein clearance.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Epidermal cells obtained from PD patients with the LRRK2-G2019S mutation showed a blunted response to LPS treatment compared to control cells.

“In agreement with our findings, a recent study reported that epidermal cells obtained from PD patients with the LRRK2-G2019S mutation showed a blunted response to LPS treatment compared to control cells (65).”

• cites [65] Lopez de Maturana R, Aguila JC, Sousa A, Vazquez N, Del Rio P, Aiastui A, Gorostidi A, Lopez de Munain A, Sanchez-Pernaute R, Leucine-rich repeat kinase 2 modulates cyclooxygenase 2 and the inflammatory response in idiopathic and genetic Parkinson’s disease. Neurobiol Aging 35, 1116– 1124 (2014). 24360742 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

NFAT plays a critical role in T cell differentiation and response.

“NFAT is known to play a critical role in T cell differentiation and response (30, 59).”

• cites [30] Hermann-Kleiter N, Baier G, NFAT pulls the strings during CD4+ T helper cell effector functions. Blood 115, 2989– 2997 (2010). 20103781 sourceNot checked — no full text of the source
• cites [59] Feske S, Calcium signalling in lymphocyte activation and disease. Nat Rev Immunol 7, 690– 702 (2007). 17703229 sourceNot checked — no full text of the source

May combine two claims.

Modulation of LRRK2 kinase activity can ameliorate neuroinflammation, neuronal loss, and behavioral deficits in a mouse model of synucleinopathy.

“Collectively, these results indicate that modulation of LRRK2 kinase activity can ameliorate neuroinflammation, neuronal loss, and behavioral deficits in a mouse model of synucleinopathy.”

• AssumptionRests on an assumption: HG-10-102-01 is a brain-permeable and selective LRRK2 kinase inhibitor.

Classified as probably not a substantial claim.

It remains controversial whether LRRK2 modulates NFAT.

“However, it remains controversial whether LRRK2 modulates NFAT.”

May combine two claims.

Exposure to neuron-released α-synuclein promotes microglial LRRK2 kinase activation, inducing nuclear translocation of NFATc2 to increase TNFα and IL-6 expression.

“Exposure to neuron-released α-synuclein promoted microglial LRRK2 kinase activation, inducing nuclear translocation of NFATc2 to increase TNFα and IL-6 expression.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The diagnosis was based on the initial clinical presentation with dementia followed by parkinsonism and the presence of cortical and subcortical α-synuclein/Ubiquitin-positive Lewy bodies.

“The diagnosis was based on the initial clinical presentation with dementia followed by parkinsonism and the presence of cortical and subcortical α-synuclein/Ubiquitin-positive Lewy bodies (71).”

• cites [71] McKeith IG, Galasko D, Kosaka K, Perry EK, Dickson DW, Hansen LA, Salmon DP, Lowe J, Mirra SS, Byrne EJ, Lennox G, Quinn NP, Edwardson JA, Ince PG, Bergeron C, Burns A, Miller BL, Lovestone S, Collerton D, Jansen EN, Ballard C, de Vos RA, Wilcock GK, Jellinger KA, Perry RH, Consensus guidelines for the clinical and pathologic diagnosis of dementia with Lewy bodies (DLB): report of the consortium on DLB international workshop. Neurology 47, 1113– 1124 (1996). 8909416 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

No animal or sample was excluded from the analysis.

“No animal or sample was excluded from the analysis.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Representative images were obtained from the subjects in randomly chosen area.

“Representative images were obtained from the subjects in randomly chosen area.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Blinding was performed during data collection and analysis.

“Blinding was performed during data collection and analysis.”

Probably not a substantial claim: may not pass S3 (load-bearing).

No outliers were excluded from the study.

“no outliers were excluded from the study”

Probably not a substantial claim: may not pass S3 (load-bearing).

Sample size was determined by availability and previous experience with α-syn transgenic mouse.

“Sample size was determined by availability and previous experience with α-syn transgenic mouse”

Probably not a substantial claim: may not pass S3 (load-bearing).

Data acquisition and quantification was performed by experimenters blinded to genotype with the exception of the in vitro molecular analyses.

“Data acquisition and quantification was performed by experimenters blinded to genotype with the exception of the in vitro molecular analyses.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Injections and dissections were conducted in a non-blinded fashion.

“Injections and dissections were conducted in a non-blinded fashion.”