The amyloid precursor protein and its derived fragments concomitantly contribute to the alterations of mitochondrial transport machinery in Alzheimer’s disease

doi:10.21203/rs.3.rs-3235215/v1

Download PDF

Article

The amyloid precursor protein and its derived fragments concomitantly contribute to the alterations of mitochondrial transport machinery in Alzheimer’s disease

https://doi.org/10.21203/rs.3.rs-3235215/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 27 May, 2024

Read the published version in Cell Death & Disease →

You are reading this latest preprint version

Mitochondria dysfunctions and mitophagy failure have been associated with several Alzheimer’s disease (AD) related molecular actors including amyloid beta (Aβ) and recently the amyloid precursor protein-C terminal fragments (APP-CTFs). The efficacy of the mitophagy process in neurons relies on regulated mitochondrial transport along axons involving a complex molecular machinery. The contribution of the amyloid precursor protein (APP) and its derived fragments to the mitochondrial transport machinery alterations in AD have not been investigated before. We report herein a change of the expression of mitochondrial transport proteins (SNPH and Miro1), motor adaptors (TRANK1 and TRAK2) and components of the dynein and kinesin motors (i.e. IC1,2 and Kif5 (A, B, C) isoforms) by endogenous APP and by overexpression of APP carrying the familial Swedish mutation (APPswe). We show that APP-CTFs and Aβ concomitantly regulate the expression of a set of transport proteins in APPswe cells treated with β- and γ-secretase inhibitors and by using cells Knock-down for presenilin 1 and 2. We further reported the impact of APP-CTFs on the expression of transport proteins in AAV-injected C99 mice brains. Our data also indicate that both APP-CTFs and Aβ oligomers (Aβo) impair the localization of transport proteins in mitochondria as demonstrated in differentiated SH-SY5Y-APPswe cells treated with β- and γ-secretase inhibitors and in Aβo-treated differentiated SH-SY5Y naive cells. Importantly, we uncover that the expression of a set of transport proteins is modulated in a disease-dependent manner in 3xTgAD mice and in human sporadic AD brains. This study highlights molecular mechanisms underlying mitochondrial transport defect in AD that likely contribute to mitophagy failure and disease progression.

Alzheimer’s disease. APP. Aβ. APP-CTFs. Mitochondria transport. Presenilins 1 and 2

Mitochondria ensures several vital functions, including energy production through the synthesis of adenosine tri-phosphate (ATP) by the oxidative phosphorylation (OXPHOS) system (1). Mitochondria organelles follow a dynamic transport via microtubules (MT) to fulfil their role locally in polarized cells such as neurons (2). Thus, the anterograde transport occurring from the nucleus to the cell’s periphery (i.e. axons, dendrites and synapses) distributes newly synthetized and healthy mitochondria, thereby ensuring the replenishment of these high energy demanding sites with ATP that is necessary for synaptic transmission and plasticity (3, 4). In an opposite manner, dysfunctional mitochondria undergo a retrograde transport backward to the soma where they are degraded by mitophagy (i.e the specific degradation systems of altered mitochondria) upon fusion of the mitophagosomes with lysosomes (5).

Mitochondrial transport along MT is achieved by a set of ATP-driven motors including kinesin for anterograde transport and cytoplasmic dynein for retrograde transport (6). Mammalian kinesin isoforms includes the neuronal Kif5A and Kif5C and the ubiquitous Kif5B (7). Cytoplasmic dynein is a multi-subunit nanomotor composed of intermediate chains (ICs), light intermediate chains (LICs) and light chains (LCs) built around a core of large dimeric heavy chains (HCs), which perform ATP hydrolysis (8). These motor proteins can carry several organelles and vesicles and interact with specific cargoes or cargo-adaptor proteins to transport mitochondria (6). Among motor adaptors proteins, kinesin-binding proteins (TRAK1 and TRAK2) are implicated in the trafficking of different organelles, including endosomes and are linked indirectly to mitochondria through the interaction with mitochondrial rho (MIRO), a RHO family GTPase expressed at the mitochondrial outer membrane (9). Miro1 isoform is the major mitochondrial acceptor in neurons (10). The mechanisms linking dynein to mitochondria are not well characterized. However, it seems that MIRO may also be required for dynein-mediated retrograde mitochondrial transport (11). Another protein that regulates mitochondrial transport is syntaphilin (SNPH) that blocks the global movement of mitochondria by directly anchoring the organelle on MT (4).

Alzheimer’s disease (AD) is a neurodegenerative disease initially defined by a proteinopathy associated with the accumulation of amyloid beta (Aβ) peptides that are produced through the sequential cleavage of the amyloid precursor protein (APP) by the β- and the γ-secretases (12). These Aβ peptides accumulate extracellularly in senile plaques (13) and interact with different intracellular cellular components and organelles, including mitochondria (14, 15). Besides Aβ, APP processing generates other fragments that have several toxic features contributing to the etiology of AD (16), namely the APP C-terminal fragments (APP-CTFs: C99 and C83). C99 fragment results from of the cleavage of APP by β- secretase and C83 fragment derives from the cleavage of both APP and C99 by α-secretase (17–20). Accumulation of APP-CTFs in cellular models mimicking familial and sporadic forms of AD triggers an autolysosomal defect, which in turn impair APP-CTFs degradation and induce their pathogenic accumulation (21, 22). In a recent study, we reported that APP-CTFs accumulation is associated with mitochondrial structure and function alterations and triggers mitophagy failure in both cellular and mice AD models (23). These findings are supported by a recent study demonstrating the accumulation of APP-CTFs in mitochondria and their causality of impaired mitophagy function in AD patient-induced neural stem cells (iNSCs) (24). Importantly, APP-CTFs accumulation in sporadic human AD brains correlates with a mitophagy failure molecular signature, and is associated with neurodegeneration (23, 25).

In parallel to mitochondria structure, function and mitophagy alterations, isolated studies reported impaired mitochondrial transport in AD-related conditions (26). However, the molecular mechanisms underlying mitochondrial transport defect in AD remain partially understood. Here, we investigated the specific implication of APP, APP-CTFs and Aβ on the expression and localization of mitochondrial transport components (i.e. SNPH, Miro1, TRAK1 and 2, Kif5 A, B, C isoforms and IC 1, 2) using complementary cellular and mice AD models. We also studied the molecular signature of mitochondrial transport proteins associated with AD progression in post-mortem human brains.

Human brain samples

We studied control and sporadic AD (SAD) post-mortem human brains (Control, n = 7; I-III, n = 5; and IV-VI, n = 14). Cases were anonymized, but information was provided regarding sex, age at death, and neuropathology (Table 1). Studies involving human brain samples were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical studies. Brain samples were obtained from the Brain Bank “NeuroCEB” run by a consortium of Patients Associations: ARSEP (association for research on multiple sclerosis), CSC (cerebellar ataxias), and France Parkinson. The consents were signed by the patients themselves or their next of kin in their name, in accordance with the French Bioethical (agreement AC-2013-1887).

Table 1

Demographic data and neuropathology evolution of human brain samples described by Braak neurofibrillary stages and Thal amyloid stages, used in SDS-PAGE. Controls (CT) are brain samples isolated from post-mortem patients diagnosed as negative for several neuropathologies; we also used control post- mortem brains samples diagnosed as negative for AD pathology and with amyotrophic lateral sclerosis (ALS); NA: not available; PMD: Post-mortem delay in hours; T1 : region of the temporal lobe.
	Thal stage	Braak stage	Gender	Age	PMD (h)	Brain area
CT	-	-	H	71	26	T1
CT	-	-	H	84	32	T1
CT	-	-	F	71	15	T1
ALS	-	-	F	62	44	T1
ALS	-	-	H	55	21	T1
CT	-	-	F	89	12	T1
CT	-	-	F	52	29	T1
AD	NA	I-II	F	68	51	T1
AD	0	II	F	83	21	T1
AD	2	II	M	85	10	T1
AD	0	III	F	92	NA	T1
AD	0	III	M	70	31	T1
AD	2	IV	F	76	28	T1
AD	NA	IV	M	92	NA	T1
AD	NA	V	M	71	NA	T1
AD	5	V	M	82	25	T1
AD	NA	V	F	80	51	T1
AD	0	VI	F	78	18	T1
AD	5	VI	F	65	41	T1
AD	4–5	VI	F	75	7	T1
AD	4	VI	F	89	26	T1
AD	NA	VI	F	93	21	T1
AD	5	VI	F	91	34	T1
AD	5	VI	F	55	58	T1
AD	NA	VI	H	67	30	T1
AD	1	VI	F	82	NA	T1

Animals

Experiments were performed on hippocampi dissected from 3xTgAD mice (APPswe: KM670/671NL, TauP301L, and presenelin1 (PS1) M146V) (n = 8 aged 4 months and n = 7 aged 13 months) (27) and wild-type mice (WT; non-transgenic) (n = 7 aged 4 months and n = 5 aged 13 months). We also used brains of male mice aged 6 months and injected at day 1 post-natal with adeno-associated-virus (AAV)-C99 (n = 5) or AAV-Free (empty virus) (n = 4) (22). Mice were housed with a 12:12 h light/dark cycle and were given free access to food and water.

Ethical Approval

Studies on animals were performed following the guidelines established by the European community council (Directive of November 24^th, 1986), and applying ethical standards of the institution. Animal experimentation protocols were approved by Nice university Animal care and use Committee, and the National Council on animal care of the French Ministry of Higher Education and Research (project #00253.02 and APAFIS#20495-201904231352370). These projects were evaluated and followed by the welfare animal use committee of the institute.

Cells

We used human SH-SY5Y neuroblastoma cells (CRL-2266, ATCC) stably expressing empty pcDNA3.1 vector (Control), or the human βAPP harboring the double Swedish mutations cDNA (APPswe: APPKM670/671NL) (28). We also used mouse embryonic fibroblasts: MEFs either control (MEF APPWT or MEF PSWT), invalidated for APP and for APP-like proteins 1 and 2 (APLP1 and APLP2) (MEF APPKO) (29), or doubly knocked out for presenilin 1 (PS1) and presenilin 2 (PS2) (PSDKO) (30). Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, GIBCO 41965-039) supplemented with 10% Fetal Bovine Serum (FBS), penicillin (100 U/ml) and streptomycin (50 μg/ml) (Pen/Strep), and incubated at 37°C in 5% CO2 atmosphere. Control and APPswe SHSY5Y cells were cultured in the presence of 400 μg/ml geneticin (MP Biomedicals) (28).

Treatments

Cells were treated with DMSO (Sigma D2650) as control (vehicle), γ-secretase inhibitor ELDN006 (3nM in DMSO) (31) or β-secretase inhibitor LY2811376 (30nM in DMSO) (32) for 16 hours. Differentiated SHSY-5Y-APPswe were treated with γ-secretase inhibitor (0.3nM) for 16 hours to limit cell death.

Differentiation of SHSY5Y cell lines

We used a protocol adapted from a previously reported protocol (33). Briefly, cells were plated in a 6-well plate at a confluence of 30-40% to reach 70% maximum 5 days later. The first week of culture, we used M1 medium (DMEM supplemented with Pen/Strep and 2.5% of FBS, and 10µM retinoic acid (RA, Merck R2526). Cells were then split into new 6-well plates and cultured in M2 medium (DMEM supplemented with Pen/Strep, 1% of FBS, and 10µM RA). Cells were then split two days later in plates coated with 0.5µg/ml Poly-L-lysin (Sigma-Aldrich P2636) in neurobasal medium complemented with B27 (1X), Pen/Strep, 50ng/mL BNDF (Merck B3795), 2mM Glutamax (Gibco 35050061), 20mM KCL solution and 10µM RA. Differentiated cells were used 3 weeks later.

Aβ production

Dry Aβ 1-42 (Bachem AG 4014447.1000) (1mg) was diluted in HFIP (1, 1, 1, 3, 3, 3, 3 – Hexafluoro-2-propanol) (SIGMA 105228-5G) for 1 hour to obtain a 1mM homogenous solution, then aliquoted into microfuge tubes and lyophilized under laminar hood overnight (O/N) to ensure complete evaporation. To form Aβ oligomers, peptide films were resuspended to 5mM in DMSO, diluted in ice cold PBS. Aggregation was allowed to proceed for 24 hours at 4°C before the peptide solution was centrifuged at 14000 x g for 10 minutes at 4°C. Supernatant containing Oligomeric Aβ (Aβo) was used at ≈ 5µM for 16 hours.

Immunofluorescence

Differentiated control or APPswe-expressing SH-SY5Y cells were fixed with 4% paraformaldehyde (PFA) for 10 minutes, washed with 1X PBS, and then permeabilized with 0.5% Triton for 5 minutes. Non-specific binding sites were blocked for 1hour in 3% PBS-BSA containing 0.005% tween at room temperature. Primary antibodies were diluted (Supplement table 1) in PBS-BSA 0.3% containing 0.0005% tween and applied overnight (ON). After three washes with 1xPBS, secondary antibodies were applied for one hour. Nuclei were stained with DAPI (1/5000, Invitrogen) and slides were mounted with Vectamount medium (Vector). Images were taken with a confocal Leica TCS SP5 microscope or LSM 780 and processed using ImageJ software.

Protein extraction and SDS-PAGE and western blot analysis

Total protein extracts were obtained from cells, dissected mice hippocampi of WT and 3xTgAD, cortices of AAV-Free or AAV-C99 mice, or human temporal cortex using lysis buffer (50 mM Tris pH 8, 10% glycerol, 200mM NaCl, 0.5% Nonidet p-40, and 0.1 mM EDTA) supplemented with protease inhibitors (Complete cocktail, Roche diagnostics 04693132001). Proteins were resolved by SDS-PAGE following standard procedures and APP-CTFs were resolved on 16.5% Tris-Tricine SDS-PAGE (23). Information about the primary antibodies used for western blot can be found in Supplement table 1.

Statistical analyses

Data were expressed as means ± SEM. Sample size for each experiment is indicated in the Figure captions. The quantification of imaging experiments was performed on different (> 5) fields of view in at least three independent experiments. Data were analyzed with GraphPad Prism version 9 for Windows (GraphPad Software, La Jolla, CA, USA; https://www.graph pad.com). Data were first analyzed for normal distribution. We used the Mann–Whitney test when the two groups of variables have not passed the normality test. Groups of more than two variables that have passed normality test were analyzed by one-way ANOVA with Dunnett’s or Tukey’s multiple comparisons post-test. Kruskal–Wallis test and Dunn’s multiple comparisons post-test was used when groups of variables have not passed the normality test. Correlations were analyzed with Spearman’s coefficient. Significant differences are: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and ns: non-significant.

APP and its derived fragments modulate the expression of proteins of the mitochondrial transport machinery

We studied the expression of the mitochondrial stop protein syntaphilin (SNPH), the mitochondrial rho protein (Miro1), the kinesin-binding proteins (TRAK1 and TRAK2), the motor kinesin (Kif5 (A, B, C) isoforms), and the intermediate chains (ICs) of the cytoplasmic dynein motor (lC1,2) in neuroblastoma cells expressing APP carrying the double Swedish familial mutations KM670/671NL (APPswe) and their control cells expressing pcDNA3.1 empty vector (Control) (28) (Fig.1). This cellular model has been previously described to accumulate amyloid precursor protein (APP)-derived fragments (i.e. Aβ and APP-CTFs) and to manifest mitochondrial structure and function alterations and a defect of mitophagy process (23). We noticed a reduction in the expression of Miro1 and TRAK1 (Fig., 1a, c), Kif5 (A, B, C) (Fig. 1a, d), and an increase in IC1, 2 level (Fig., 1a, d) in APPswe versus control cells, while the expression of SNPH (Fig., 1a) and TRAK2 (Fig., 1c) remained unchanged. We identified a low molecular weight (» 50 kDa) second band recognized by the Kif5 (A, B, C) antibody that we labelled as Kif5 (low) showing in APPswe cells a mirror upregulation versus full-length Kif5 (A, B, C) (Fig. 1a, d).

Because APP is involved in several housekeeping cellular functions (34), we investigated whether the down expression of full-length wild-type APP may impact the expression of mitochondrial transport proteins. Since APP shares some functional redundancies with the amyloid precursor-like proteins APLP1 and APLP2, we compared control mouse embryonic fibroblasts (MEF APPWT) with cells depleted of APP and of APP-like proteins 1 and 2 (APLP1 and APLP2) (MEF APPKO). We confirmed the absence of APP expression and observed a reduced endogenous APP-CTFs levels in MEFs APPKO cells (Fig. 2a). We evidenced an enhanced expression of SNPH, Miro1, TRAK1, TRAK2, and Kif5 (A, B, C) in MEFs APPKO cells when compared to controls (Fig. 2a, b, c, d) and noticed a mirror downregulation of Kif5 (low) band versus full-length Kif5 protein and of IC1, 2 protein in MEFs APPKO (Fig., 1a, d).

Neuroblastoma cells show a “compacted dense shape” of the mitochondria reticulum that was technically not adapted for the analyses of mitochondrial movement using imagery and time-laps acquisition. Thus, we studied mitochondria movement in MEF presenting “a flat shape mitochondria reticulum” and we revealed a reduced mitochondrial movement in MEF APPKO compared to MEF APPWT (Fig. 2e, and supplement Video. 1). Since the expression levels of several proteins implicated in mitochondria movement are upregulated in MEF APPKO, we speculate that the reduction of mitochondria motile fraction in APPKO cells is associated with enhanced expression of SNPH known to block mitochondria movement (Fig. 2a, b).

Together, these pieces of data point-out a contribution of endogenous APP and of APP-derived fragments accumulation in the control of the expression of a set of proteins constituting the mitochondrial transport machinery complex.

Accumulation of both Aβ and APP-CTFs modulates the expression of several proteins of the mitochondrial transport machinery complex

In order to discriminate between the impact of Aβ and APP-CTFs on the expression of transport proteins, we used: i) the β- and the g-secretases inhibitors in APPswe cells, to modulate the processing of APP and the levels of APP-CTFs (C99 and C83) and Aβ; and ii) MEF double knock-out for presenilins 1 and 2 (components of the g-secretase complex) (MEF PSDKO). We previously reported that both β- and g-secretases inhibitors block Aβ production in APPswe cells (35). We also performed a new set of experiments and confirmed that, while the inhibition of the g-secretase triggers an accumulation of both C99 and C83 fragments in APPswe cells (Fig. 3a, and Supplement Fig. 1a), the inhibition of the β-secretase blocks C99 production and enhances the level of C83 (Supplement Fig. 1a). We then showed that g-secretase inhibition in APPswe cells triggers a reduction in SNPH levels (Fig. 3b, c) and potentiates the reduction in the expression of KIF5 (A, B, C) (Fig. 3b, e). In parallel, we observed that the reduced expression in Miro1 and TRAK1 in APPswe cells remained unchanged upon g-secretase inhibition (Fig. 3b, d), suggesting the prevalence of APP-CTFs accumulation over Aβ blockade in these alterations. Finally, we showed that the inhibition of the g-secretase abolishes the APP-induced increases in Kif5 (low) and IC1,2 (Fig. 2b, f), thus signifying a potential implication of Aβ accumulation in these alterations. These observations indicate that the expression levels of mitochondrial transport proteins are mitigated by either APP-CTFs or Aβ accumulation.

In the same sets of experiments, we assessed the impact of g-secretase inhibition in control cells and noticed that the endogenous accumulation of APP-CTFs (23) does not impact the expression of mitochondrial transport proteins (supplement Fig. 2 a-c). We also reported that β-secretase inhibition in APPswe cells does not impact the expression of our proteins of interest (Supplement Fig. 1), indicating the prevalence of C99 over C83 in the observed effects upon g-secretase inhibition.

Since g-secretase has over 150 substrates (36), we undertook to ascertain that the changes we observed in APPswe cells are genuinely linked to APP-CTFs over accumulation and not to other g-secretase substrates. We thus demonstrated that the reported modulations of mitochondrial transport proteins (Fig. 2) are not observed in MEF APPKO treated with the g-secretase inhibitor (supplement Fig. 3). This confirmed that the modulation of protein expressions observed in APPswe cells were not due to the processing of other substrates than APP by g-secretase.

In addition, we studied the expression of the mitochondrial transport machinery complex in MEF PSDKO in which an accumulation of APP-CTFs occurs (Fig. 4a). Interestingly, MEF PSDKO showed a significant reduction in SNPH, Miro1, TRAK1, TRAK2 and IC1,2 levels when compared to control MEF (MEFs PSWT) (Fig. 4a, b, c, d), thus almost mimicking the results obtained in APPswe cells treated with the g-secretase inhibitor and supporting a contribution of APP-CTFs accumulation to the alteration of the expression of mitochondrial transport machinery.

Intriguingly, we noticed that the expression of full-length Kif5 (A, B, C) and its lower band were not altered in MEF PSDKO (Fig. 4a, d), while they were regulated in APPswe-cells treated with the g-secretase inhibitor. This may suggest that Kif5 low band production is regulated in a cell-type specific manner (SH-SY5Y versus MEFs) and/or does not solely implicates APP-CTFs accumulation.

All together, these data point-out a drastic deregulation of mitochondrial transport proteins linked to PS knock-down. In fact, we revealed a reduced mitochondrial movement in MEF PSDKO when compared to their controls (Fig. 4e, and supplement Video 2).

Thus, both genetic depletion and pharmacological blockade of g-secretase concur to support a major deleterious effect of APP-CTFs accumulation on the expression of mitochondrial transport machinery.

APP and some of its derived fragments alter the colocalization of several proteins of the mitochondrial transport machinery complex with mitochondria

We used differentiated SH-SY5Y cellular models to investigate the localization of proteins of the mitochondrial transport machinery complex in a polarized cellular context. We first notice a reduction of the proliferation of control and APPswe cells and validate their morphological changes towards a phenotype of polarized cells approaching that of neurons (Fig. 5a). Differentiated control and APPswe SH-SY5Y cells show branched morphology as revealed by an increase of the staining with two microtubule markers (β3-tubulin and microtubule-associated proteins, MAP2) (Fig. 5a). Differentiated cells also show an increase of the staining of neuronal marker NeuN and of tyrosine hydroxylase (a marker for dopaminergic and adrenergic neurons) (Fig. 5a). We also verified that differentiated APPswe cells provides an AD “neuronal like” study model accumulating APP-CTFs upon g-secretase inhibitor treatment (Supplement Fig. 4).

We then determined the intracellular localization of SNPH, Miro1, TRAK1, TRAK2, Kif5 and IC1,2 by immunofluorescence and we stained mitochondria using TOMM20 antibody. Differentiated APPswe cells show a reduction of the localization of SNPH (Fig. 5b, c), and Kif5 (Fig. 5b, e) in mitochondria, and an increase of the localization of TRAK1 (Fig. 5b, d) in mitochondria, while the localization of Miro1, TRAK2 and IC1,2 remained unchanged (Fig. 5c-e). Interestingly, g-secretase inhibition amplifies this phenomenon by triggering an additional diminution of the mitochondrial localization of SNPH and Miro1 (Fig. 6a, b), and of TRAK1 and TRAK2 (Fig. 6a, c). These data demonstrate that the localization of a set of transport machinery components to mitochondria is impacted by APP-CTFs accumulation. Noticeably, we observed an increase of the localization of Kif5 in mitochondria upon g-secretase inhibition (Fig. 6a, d), while the localization of IC1,2 remained unchanged (Fig. 6d). It should be mentioned here that Kif5 (A, B, C) antibody used in the immunofluorescence analyses cannot discriminate between full-length Kif5 or cleaved Kif5 (low) forms as detected by SDS-PAGE (Fig. 1-4). Intriguingly, while the expression of transport proteins was almost not altered in APPswe cells treated with β-secretase inhibitor (Supplement Fig. 1), we noticed in these cells a reduction in the localization of Miro1, TRAK 1 and TRAK2 (Supplement Fig. 5 a-c) and an increase in IC1, 2 localization to the mitochondria (Supplement Fig. 5 a, d). These data support a contribution of the a-secretase-derived APP fragment (C83) accumulation to the alteration of the localization of some transport machinery components to mitochondria. In a complementary set of experiments, we investigated the specific impact of Aβ oligomers (Aβo) on the localization of the same set of transport proteins in mitochondria using control differentiated SH-SY5Y cells. Aβo treatment triggers a reduction in the localization of SNPH, Miro1 (Fig. 6e, f) and TRAK1 (Fig. 6e, g) to mitochondria, while the localization of TRAK2, Kif5, and IC1, 2 remains unchanged (Fig. 6g, h).

Together, these data (Fig. 5 and 6) firmly demonstrate a defect of the mitochondrial transport machinery localization to mitochondria linked to both Aβ and APP-CTFs (C99 and C83).

The expression of several proteins of the mitochondrial transport machinery complex is altered in 3xTgAD and AAV-C99 mice

We then studied the expression of our proteins of interest in the hippocampus of the 3xTgAD mice that harbour an early and progressive accumulation of APP-CTFs starting at 2-3 months of age followed by the production and the accumulation of Aβ at 6 months of age and forming the amyloid plaques at 10-11 months of age (18, 27, 37). We first revealed an increase in SNPH expression in young 3xTgAD mice (aged 4 months) hippocampi when compared to age-matched control mice hippocampi (Fig. 7a, b), while the levels of the expression of Miro1, TRAK1, TRAK2, Kif5 and IC1,2 expressions remain unchanged or slightly but not significantly increased (Fig. 7a, c, d). On the contrary, we reported a significant decrease in SNPH, Miro1 and TRAK2 in old 3xTgAD mice (aged 13 months) when compared to age-matched control mice (Fig. 7e, f). In parallel, while we noticed a trend decrease of the expression of TRAK1and IC1,2 in old 3xTgAD mice hippocampi, Kif5 (A, B, C) and Kif5 (low) remain unchanged (Fig. 7a, g, h). In addition to APPswe mutation, the 3xTgAD mice overexpress a mutated form of the Tau protein (TauP301L) and are knocked-in for presenilin1 (PS1) carrying M146V mutation. Thus, in addition to an impact of APP-derived fragments, we cannot exclude a potential contribution of mutated PS1 and/or phosphorylated Tau to the alterations of the expression of the mitochondrial transport machinery complex observed in this model. To investigate the specific impact of the C99 accumulation on mitochondrial transport machinery in vivo, we used AAV-C99 mice overexpressing C99 fragment in an endogenous APP, PS and Tau background (23). Interestingly, SDS-PAGE analyses revealed a drastic reduction in the expression of SNPH, Miro1, TRAK1, TRAK2, and Kif5 (A, B, C) in AAV-C99 injected mice brains when compared to AAV-Free injected controls (Fig. 8a-d), thus revealing a role of APP-CTFs accumulation in mitochondrial transport alteration in vivo.

The expression of several proteins of the mitochondrial transport machinery complex is altered in sporadic AD human brains

We lastly questioned whether human SAD brains manifest an altered expression of mitochondrial transport proteins. We stratified AD brains into early Braak stages (I-III) and late Braak stages (IV-VI) (patients’ information in Table 1). Quantitative analyses revealed a significant reduction in the expression of SNPH (Fig. 9a, b) and TRAK2 (Fig. 8a, e) in Braak IV-VI AD brains versus age-matched non-demented control brains while TRAK2 (Fig. 9a, e) and IC1,2 (Fig. 9a, h) levels were gradually reduced in a disease-dependent manner (i.e. Braak IV-VI AD brains versus Braak stages (I-III) AD brains). In parallel, we noticed that the expressions of Miro1 and TRAK1 are slightly but not significantly reduced in Braak IV-VI AD. Interestingly, correlations studies including control and AD brains revealed a statistically significant positive correlation between the expression levels of several transport proteins (Fig. 9 i), namely between SNPH and Miro1, TRAK1, and TRAK2. Miro1 expression also positively correlates with that of TRAK1, TRAK2 and Kif5 low. Both TRAK1 and TRAK2 expressions correlate with Kif5 (A, B, C). Finally, IC1, 2 expression level correlates with TRAK2 and Kif5 (A, B, C) (Fig. 9i). These correlations have to be put in perspective with the fact that transport proteins follow a decreased expression trend in a disease-dependent manner in AD. We further documented the relationship between the level of APP-CTFs and Aβ (Fig. 9j, k and supplement Fig. 5), and the expression of transport proteins and showed statistically significant negative correlations of SNPH, and TRAK1 levels with APP-CTFs levels, and of SNPH and TRAK2 expressions with Aβ level (Fig.9 k). These AD brains also manifest enhanced phosphorylation of Tau on serine231(pTauT231) and threonine422 (pTauS422) residues quantified versus total Tau protein (tTau) (Supplement Fig. 6a). However, we found that the correlation extent is restricted to SNPH and pTauS422 but not with pTauT231 (Supplement Fig. 6b).

Together, these data support the findings in cellular and mice AD models mimicking familial forms of the disease and point-out altered expression of mitochondrial transport proteins in human AD brains, mostly contributed by APP-CTFs and Aβ.

In the current study, we unraveled several alterations in the expression and localization of different components of the mitochondrial transport machinery in cellular and mice AD models and in human post-mortem SAD brains. We reported that depletion of the endogenous full-length APP is associated with a reduced mitochondria movement that is most likely linked to an enhanced expression of the stop protein SNPH and to reduced expression of dynein (lC 1,2) motor. Moreover, the enhanced expression of Miro1, TRAK1, TRAK2, and Kif5 (A, B, C) in APPKO cells further highlight a potential direct or indirect role for the endogenous APP in the control of mitochondrial transport proteins. In fact, previous studies have shown that vesicular APP is transported along axons and that APP directly interacts with Disrupted-In-SChizophrenia 1 (DISC1), shown to regulate anterograde mitochondrial transport and to interact with Miro1 and TRAK1/2 (38). In addition, APP also interacts with the kinesin light chain subunit KLC1 (Kinesin light chain 1) and acts as a conventional cargo for kinesin-I microtubule motor protein (39).

We carried out experiments to investigate the specific impact of the accumulation of APP-derived fragments on mitochondrial transport components and used a cellular model overexpressing an AD familial mutation (SH-SY5Y APPswe). This cellular model combined with pharmacological modulation of β- and g-secretase activities allows us to demonstrate that both Aβ and C99 contribute to the alteration of the expression of mitochondrial transport components. As a matter of fact, the reduction in the expression of Miro1 and TRAK1 in APPswe cells is maintained upon g-secretase inhibition that blocks Aβ production and exacerbates APP-CTFs overaccumulation. Moreover, we reported a reduction in the expression of SNPH and of full-length Kif5 (A, B, C) in APPswe cells treated with the g-secretase inhibitor. Our results in AAV-C99 mice brains further support a pathogenic molecular signature of the mitochondrial transport proteins linked to C99 accumulation in vivo.

The defect of mitochondrial movement and the alterations of the expression of transport proteins in PSDKO cells could likely be associated to APP-CTFs accumulation in this model. However, we must not exclude that the defects reported in this model could occur in a g-secretase-independent manner, through PS-mediated control of vesicular transport and trafficking (40). This assumption is supported by a pioneer study by Kamal et al. demonstrating that vesicular PS1 is transported with APP along axons by kinesin 1 (39). On the light of our findings and this observation, one may speculate that PS depletion (or loss of function linked to familial forms of AD) could destabilize vesicular transport and impact global transport machinery.

Our experiments in APPswe cells revealed an increase of the expression of Kif5 (low band) and IC 1,2 that seems to be linked with Aβ production, since these alterations are reversed by g-secretase inhibition. Consistent with a specific toxic effect of Aβ on transport machinery, a recent study reported a reduction of Miro1 level by Aβo (41). Accordingly, we reported a reduction of Miro1 colocalization with mitochondria in differentiated neuroblastoma cells treated with Aβo.

Previous studies have suggested that APP axonal transport by kinesin-I may interfere with vesicular APP processing to Aβ and APP-CTFs occurring in an axonal membrane compartment containing β-secretase and PS1 (39). A second study demonstrated that knock-down of DISC1, is accompanied by an increase in APP-CTFα and a decrease in Aβ42 and Aβ40 levels (42). Our findings provide original observations demonstrating that endogenous full-length APP as well as the accumulation of APP-derived fragments occurring under AD conditions interfere in turn with transport machinery. Since these alterations affect both dynein and kinesin motors and implicate several mitochondrial adaptors as well as the stop protein SNPH, we may hypothesize that APP and its derived fragments impact global anterograde and retrograde mitochondrial transport under physiological and AD conditions.

Although differentiated neuroblastoma cells are not fully functional neurons, they are an alternative study system mimicking polarized primary culture of neurons. Moreover, differentiated SH-SY5Y APPswe cells still overproduce APP-derived fragments, the accumulation of which is enhanced by γ-secretase inhibitor. Using these models, we demonstrated that the loss of the localization of the transport proteins to mitochondria is an additional molecular event regulated by APP-derived fragments (C99, C83 and Aβ) and likely perturbing mitochondria anterograde and retrograde transport. These data fully consolidate our biochemical data demonstrating an alteration of the expression of a set of mitochondrial transport machinery components linked to APPswe overexpression and concomitantly implicating C99 and Aβ accumulation.

Mitophagy failure is commonly associated with several AD-related proteins (Tau, Aβ, APP-CTFs and APOE) (43) and is likely associated with a defect of mitochondrial transport along axons. Importantly, genetic deletion of Miro1 or the blockade of Miro1 ubiquitination and subsequent degradation lead to a delayed translocation of the E3 ubiquitin ligase Parkin onto damaged mitochondria and reduced mitochondrial clearance by mitophagy in both fibroblasts and cultured neurons (44). On the other side, Pink1 and Parkin act in concert to target Miro1 to proteasomal degradation to arrest mitochondria motility (45, 46). These observations and other studies have proposed Miro1 as a molecular marker for the risk of Parkinson disease and as a potential therapeutic target for the disease (47, 48). Interestingly, our data revealed that Miro1 expression is drastically reduced in APPswe cells and is also modulated in AAV-C99 and 3xTgAD aged mice brains. We also revealed that Miro1 localization to mitochondria is impacted by APP-CTFs accumulation and Aβo treatment in differentiated neuroblastoma cells. These observations place Miro1 as a key molecular actor likely controlling both mitochondrial transport and mitophagy defects occurring in AD.

We reported that SNPH expression is reduced in concert with APP-CTFs accumulation in APPswe cells, in aged 3xTgAD mice (13-month-old) hippocampi and AAV-C99 injected mice brains and in human SAD brains at advanced stages of the disease. Moreover, the reduced expression level of SNPH in human brains significantly correlates with APP-CTFs and Aβ and to a lesser extent with pathogenic pTau (S231 and T422). SNPH deficit may likely contribute to AD development or progression. In fact, SNPH is linked to synaptic dysfunction accompanying the loss of static mitochondria (4). On the contrary, enhanced level of SNPH in young 3xTgAD mice could be taken as a compensatory mechanism that ensures local mitochondria movement arrest allowing calcium buffering and ATP release at the level of altered synapses (49). In fact, previous studies reported that SNPH expression increases in mature neurons in mice (50) and play an essential role in synaptic plasticity and in the stimulation of the liberation of synaptic vesicles (51, 52).

TRAK1/2 adaptors coordinate opposing motors (i.e kinesin-1 and dynein) for the proper transport of mitochondria (6). While a previous study has shown that TRAK2 protein level is reduced by caspase3-truncated Tau (53), the specific impact of Aβ and APP-CTFs was unknown. We unraveled herein a reduction of the expression and the localization in mitochondria of TRAK1 and/or TRAK2 linked to APP-CTFs accumulation and/or Aβo treatment. Importantly, our results in human brains at late AD stages support a reduction of TRAK1/2 expression linked to AD progression that correlates with Aβ (TRAK2) or APP-CTFs (TRAK1).

The contribution of the kinesin Kif5 (A, B, C) to mitochondria transport defect in AD is still not clear. While a previous study showed enhanced Kif5A levels at late Braak stages (54), a second paper reported a diminution of Kif5A by Aβ (55). Our results show on the one hand an Aβ-dependent Kif5 (A, B, C) down-expression in APPswe cells and on the other hand that the lesser localization of KiF5 (A, B, C) to mitochondria is linked to both APP-CTFs and Aβ. Strikingly, we did not observe any significant alteration of the expression of Kif5 (A, B, C) or its cleaved-form in human post-mortem brains cohort. Moreover, while IC1, 2 down expression positively correlates with TRAK2 and Kif5 (A, B, C) levels, it did not correlate with any of AD related proteins. These observations may question the genuine role of Kif5 (A, B, C) and IC1,2 in AD development.

Together, our data unravel novel molecular mechanisms impacting mitochondria transport, the defect of which may contribute to the loss of the transport of dysfunctional mitochondria to their sites for degradation by mitophagy and of healthy ones for the delivery of ATP to synapses. This study will certainly stimulate further research targeting mitochondria transport machinery as a mean to modulate simultaneously defects of mitochondria movement and mitophagy processes occurring in AD.

Acknowledgements

We acknowledge PhD fellow support from research ministry to LVB. We thank Dr LaFerla for providing 3xTgAD mice, Dr. Fraser for the APP-Cter antibody and Dr. U. Muller for MEF APPKO and their control MEF and Dr. De Strooper for MEF PSDKO and their control MEF.

Funding

This work was supported by LABEX (excellence laboratory, program investment for the future) DISTALZ (Development of Innovative Strategies for a Transdisciplinary approach to Alzheimer’s disease) (FC and LB), IDEX UCAJedi: Appel Jeunes Chercheurs 2022 (MC and FE), France Alzheimer (Grant # 8217) (MC), Université Côte d’Azur (Living-systems-complexity-and-diversity) (MC), and São Paulo Research Foundation (FAPESP, Grant #2018/14289-6) (PFK and MC).

Author Contributions

MC conceived and designed the study, analyzed data and wrote the manuscript. LVB set-up and performed experiments, analyzed data and participated to manuscript writing. FE, BD, PFK conducted biochemistry in mice models. RPP produced AAV-C99 mice. C. Bauer maintained mice colonies. SE and LB performed Tau ELISA analyses.

Conflict of Interests

Authors declare they have no financial interests.

Consent for publication

All authors read the final version of the article and consent to its publication.

Data Availability

The manuscript did not generate data sets.

Kann O, Kovacs R. Mitochondria and neuronal activity. Am J Physiol Cell Physiol. 2007;292(2):C641-57.
Seager R, Lee L, Henley JM, Wilkinson KA. Mechanisms and roles of mitochondrial localisation and dynamics in neuronal function. Neuronal Signal. 2020;4(2):NS20200008.
Rossi MJ, Pekkurnaz G. Powerhouse of the mind: mitochondrial plasticity at the synapse. Curr Opin Neurobiol. 2019;57:149-55.
Kang JS, Tian JH, Pan PY, Zald P, Li C, Deng C, et al. Docking of axonal mitochondria by syntaphilin controls their mobility and affects short-term facilitation. Cell. 2008;132(1):137-48.
Roney JC, Cheng XT, Sheng ZH. Neuronal endolysosomal transport and lysosomal functionality in maintaining axonostasis. J Cell Biol. 2022;221(3).
Kruppa AJ, Buss F. Motor proteins at the mitochondria-cytoskeleton interface. J Cell Sci. 2021;134(7).
Kanai Y, Okada Y, Tanaka Y, Harada A, Terada S, Hirokawa N. KIF5C, a novel neuronal kinesin enriched in motor neurons. J Neurosci. 2000;20(17):6374-84.
Pfister KK, Shah PR, Hummerich H, Russ A, Cotton J, Annuar AA, et al. Genetic analysis of the cytoplasmic dynein subunit families. PLoS Genet. 2006;2(1):e1.
Brickley K, Stephenson FA. Trafficking kinesin protein (TRAK)-mediated transport of mitochondria in axons of hippocampal neurons. J Biol Chem. 2011;286(20):18079-92.
Zinsmaier KE. Mitochondrial Miro GTPases coordinate mitochondrial and peroxisomal dynamics. Small GTPases. 2021;12(5-6):372-98.
Russo GJ, Louie K, Wellington A, Macleod GT, Hu F, Panchumarthi S, et al. Drosophila Miro is required for both anterograde and retrograde axonal mitochondrial transport. J Neurosci. 2009;29(17):5443-55.
Andrew RJ, Kellett KA, Thinakaran G, Hooper NM. A Greek Tragedy: The Growing Complexity of Alzheimer Amyloid Precursor Protein Proteolysis. J Biol Chem. 2016;291(37):19235-44.
Scheltens P, De Strooper B, Kivipelto M, Holstege H, Chetelat G, Teunissen CE, et al. Alzheimer's disease. Lancet. 2021;397(10284):1577-90.
Del Prete D, Suski JM, Oulès B, Debayle D, Gay AS, Lacas-Gervais S, et al. Localization and Processing of the Amyloid-β Protein Precursor in Mitochondria-Associated Membranes. Journal of Alzheimer's Disease. 2017;55(4):1549-70.
Eysert F, Kinoshita PF, Mary A, Vaillant-Beuchot L, Checler F, Chami M. Molecular Dysfunctions of Mitochondria-Associated Membranes (MAMs) in Alzheimer's Disease. Int J Mol Sci. 2020;21(24).
De Strooper B, Karran E. The Cellular Phase of Alzheimer's Disease. Cell. 2016;164(4):603-15.
Kim S, Sato Y, Mohan PS, Peterhoff C, Pensalfini A, Rigoglioso A, et al. Evidence that the rab5 effector APPL1 mediates APP-betaCTF-induced dysfunction of endosomes in Down syndrome and Alzheimer's disease. Mol Psychiatry. 2016;21(5):707-16.
Bourgeois A, Lauritzen I, Lorivel T, Bauer C, Checler F, Pardossi-Piquard R. Intraneuronal accumulation of C99 contributes to synaptic alterations, apathy-like behavior, and spatial learning deficits in 3xTgAD and 2xTgAD mice. Neurobiol Aging. 2018;71:21-31.
Mondragon-Rodriguez S, Gu N, Manseau F, Williams S. Alzheimer's Transgenic Model Is Characterized by Very Early Brain Network Alterations and beta-CTF Fragment Accumulation: Reversal by beta-Secretase Inhibition. Front Cell Neurosci. 2018;12:121.
Lauritzen I, Pardossi-Piquard R, Bourgeois A, Becot A, Checler F. Does Intraneuronal Accumulation of Carboxyl-terminal Fragments of the Amyloid Precursor Protein Trigger Early Neurotoxicity in Alzheimer's Disease? Curr Alzheimer Res. 2019;16(5):453-7.
Jiang Y, Mullaney KA, Peterhoff CM, Che S, Schmidt SD, Boyer-Boiteau A, et al. Alzheimer's-related endosome dysfunction in Down syndrome is Abeta-independent but requires APP and is reversed by BACE-1 inhibition. Proc Natl Acad Sci U S A. 2010;107(4):1630-5.
Lauritzen I, Pardossi-Piquard R, Bourgeois A, Pagnotta S, Biferi M-G, Barkats M, et al. Intraneuronal aggregation of the β-CTF fragment of APP (C99) induces Aβ-independent lysosomal-autophagic pathology. Acta Neuropathologica. 2016;132:257-76.
Vaillant-Beuchot L, Mary A, Pardossi-Piquard R, Bourgeois A, Lauritzen I, Eysert F, et al. Accumulation of amyloid precursor protein C-terminal fragments triggers mitochondrial structure, function, and mitophagy defects in Alzheimer's disease models and human brains. Acta Neuropathol. 2021;141(1):39-65.
Lee SE, Kwon D, Shin N, Kong D, Kim NG, Kim HY, et al. Accumulation of APP-CTF induces mitophagy dysfunction in the iNSCs model of Alzheimer's disease. Cell Death Discov. 2022;8(1):1.
Pulina MV, Hopkins M, Haroutunian V, Greengard P, Bustos V. C99 selectively accumulates in vulnerable neurons in Alzheimer's disease. Alzheimers Dement. 2020;16(2):273-82.
Flannery PJ, Trushina E. Mitochondrial dynamics and transport in Alzheimer's disease. Mol Cell Neurosci. 2019;98:109-20.
Oddo S, Caccamo A, Shepherd JD, Murphy MP, Golde TE, Kayed R, et al. Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Abeta and synaptic dysfunction. Neuron. 2003;39(3):409-21.
Oules B, Del Prete D, Greco B, Zhang X, Lauritzen I, Sevalle J, et al. Ryanodine receptor blockade reduces amyloid-beta load and memory impairments in Tg2576 mouse model of Alzheimer disease. J Neurosci. 2012;32(34):11820-34.
Heber S, Herms J, Gajic V, Hainfellner J, Aguzzi A, Rulicke T, et al. Mice with combined gene knock-outs reveal essential and partially redundant functions of amyloid precursor protein family members. J Neurosci. 2000;20(21):7951-63.
Herreman A, Serneels L, Annaert W, Collen D, Schoonjans L, De Strooper B. Total inactivation of gamma-secretase activity in presenilin-deficient embryonic stem cells. Nat Cell Biol. 2000;2(7):461-2.
Probst G, Aubele DL, Bowers S, Dressen D, Garofalo AW, Hom RK, et al. Discovery of (R)-4-cyclopropyl-7,8-difluoro-5-(4-(trifluoromethyl)phenylsulfonyl)-4,5-dihydro- 1H-pyrazolo[4,3-c]quinoline (ELND006) and (R)-4-cyclopropyl-8-fluoro-5-(6-(trifluoromethyl)pyridin-3-ylsulfonyl)-4,5-dihydr o-2H-pyrazolo[4,3-c]quinoline (ELND007): metabolically stable gamma-secretase Inhibitors that selectively inhibit the production of amyloid-beta over Notch. J Med Chem. 2013;56(13):5261-74.
May PC, Dean RA, Lowe SL, Martenyi F, Sheehan SM, Boggs LN, et al. Robust central reduction of amyloid-beta in humans with an orally available, non-peptidic beta-secretase inhibitor. J Neurosci. 2011;31(46):16507-16.
Shipley MM, Mangold CA, Szpara ML. Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line. J Vis Exp. 2016(108):53193.
Cho Y, Bae HG, Okun E, Arumugam TV, Jo DG. Physiology and pharmacology of amyloid precursor protein. Pharmacol Ther. 2022;235:108122.
Bussiere R, Lacampagne A, Reiken S, Liu X, Scheuerman V, Zalk R, et al. Amyloid beta production is regulated by beta2-adrenergic signaling-mediated post-translational modifications of the ryanodine receptor. J Biol Chem. 2017;292(24):10153-68.
Wolfe MS. Unraveling the complexity of gamma-secretase. Semin Cell Dev Biol. 2020;105:3-11.
Lauritzen I, Pardossi-Piquard R, Bauer C, Brigham E, Abraham JD, Ranaldi S, et al. The beta-secretase-derived C-terminal fragment of betaAPP, C99, but not Abeta, is a key contributor to early intraneuronal lesions in triple-transgenic mouse hippocampus. J Neurosci. 2012;32(46):16243-1655a.
Quintero H, Shiga Y, Belforte N, Alarcon-Martinez L, El Hajji S, Villafranca-Baughman D, et al. Restoration of mitochondria axonal transport by adaptor Disc1 supplementation prevents neurodegeneration and rescues visual function. Cell Rep. 2022;40(11):111324.
Kamal A, Stokin GB, Yang Z, Xia CH, Goldstein LS. Axonal transport of amyloid precursor protein is mediated by direct binding to the kinesin light chain subunit of kinesin-I. Neuron. 2000;28(2):449-59.
Vetrivel KS, Zhang YW, Xu H, Thinakaran G. Pathological and physiological functions of presenilins. Mol Neurodegener. 2006;1:4.
Kam MK, Lee DG, Kim B, Huh J-W, Lee HJ, Park Y-H, et al. Amyloid-beta oligomers induce Parkin-mediated mitophagy by reducing Miro1. The Biochemical Journal. 2020.
Shahani N, Seshadri S, Jaaro-Peled H, Ishizuka K, Hirota-Tsuyada Y, Wang Q, et al. DISC1 regulates trafficking and processing of APP and Abeta generation. Mol Psychiatry. 2015;20(7):874-9.
Mary A, Eysert F, Checler F, Chami M. Mitophagy in Alzheimer's disease: Molecular defects and therapeutic approaches. Mol Psychiatry. 2022.
Lopez-Domenech G, Howden JH, Covill-Cooke C, Morfill C, Patel JV, Burli R, et al. Loss of neuronal Miro1 disrupts mitophagy and induces hyperactivation of the integrated stress response. EMBO J. 2021;40(14):e100715.
Wang X, Winter D, Ashrafi G, Schlehe J, Wong YL, Selkoe D, et al. PINK1 and Parkin target Miro for phosphorylation and degradation to arrest mitochondrial motility. Cell. 2011;147(4):893-906.
Shlevkov E, Kramer T, Schapansky J, LaVoie MJ, Schwarz TL. Miro phosphorylation sites regulate Parkin recruitment and mitochondrial motility. Proc Natl Acad Sci U S A. 2016;113(41):E6097-E106.
Grossmann D, Berenguer-Escuder C, Chemla A, Arena G, Kruger R. The Emerging Role of RHOT1/Miro1 in the Pathogenesis of Parkinson's Disease. Front Neurol. 2020;11:587.
Hsieh CH, Li L, Vanhauwaert R, Nguyen KT, Davis MD, Bu G, et al. Miro1 Marks Parkinson's Disease Subset and Miro1 Reducer Rescues Neuron Loss in Parkinson's Models. Cell Metab. 2019;30(6):1131-40 e7.
Lin M-Y, Cheng X-T, Tammineni P, Xie Y, Zhou B, Cai Q, et al. Releasing Syntaphilin Removes Stressed Mitochondria from Axons Independent of Mitophagy under Pathophysiological Conditions. Neuron. 2017;94(3):595-610.e6.
Zhou B, Yu P, Lin MY, Sun T, Chen Y, Sheng ZH. Facilitation of axon regeneration by enhancing mitochondrial transport and rescuing energy deficits. J Cell Biol. 2016;214(1):103-19.
Lao G, Scheuss V, Gerwin CM, Su Q, Mochida S, Rettig J, et al. Syntaphilin: a syntaxin-1 clamp that controls SNARE assembly. Neuron. 2000;25(1):191-201.
Boczan J, Leenders AG, Sheng ZH. Phosphorylation of syntaphilin by cAMP-dependent protein kinase modulates its interaction with syntaxin-1 and annuls its inhibitory effect on vesicle exocytosis. J Biol Chem. 2004;279(18):18911-9.
Quintanilla RA, Tapia-Monsalves C, Vergara EH, Perez MJ, Aranguiz A. Truncated Tau Induces Mitochondrial Transport Failure Through the Impairment of TRAK2 Protein and Bioenergetics Decline in Neuronal Cells. Front Cell Neurosci. 2020;14:175.
Hares K, Miners JS, Cook AJ, Rice C, Scolding N, Love S, et al. Overexpression of Kinesin Superfamily Motor Proteins in Alzheimer's Disease. J Alzheimers Dis. 2017;60(4):1511-24.
Wang Q, Tian J, Chen H, Du H, Guo L. Amyloid beta-mediated KIF5A deficiency disrupts anterograde axonal mitochondrial movement. Neurobiol Dis. 2019;127:410-8.

(Not answered)

Download PDF

Journal Publication

published 27 May, 2024

Read the published version in Cell Death & Disease →

Editorial decision: revise
06 Dec, 2023
Review #1 received at journal
03 Oct, 2023
Reviewer #2 agreed at journal
08 Sep, 2023
Reviewer #1 agreed at journal
07 Sep, 2023
Reviewers invited by journal
04 Sep, 2023
Submission checks completed at journal
07 Aug, 2023
First submitted to journal
04 Aug, 2023
Editor assigned by journal
04 Aug, 2023

You are reading this latest preprint version

The amyloid precursor protein and its derived fragments concomitantly contribute to the alterations of mitochondrial transport machinery in Alzheimer’s disease

Status:

Journal Publication

Version 1

Abstract

Figures

Introduction

Materials and methods

Human brain samples

Animals

Results

Discussion

Declarations

References

Additional Declarations

Supplementary Files

Status:

Journal Publication

Version 1