The spectrum of neurodevelopmental, neuromuscular and neurodegenerative disorders due to defective autophagy

Primary dysfunction of autophagy due to Mendelian defects affecting core components of the autophagy machinery or closely related proteins have recently emerged as an important cause of genetic disease. This novel group of human disorders may present throughout life and comprises severe early-onset neurodevelopmental and more common adult-onset neurodegenerative disorders. Early-onset (or congenital) disorders of autophagy often share a recognizable "clinical signature," including variable combinations of neurological, neuromuscular and multisystem manifestations. Structural CNS abnormalities, cerebellar involvement, spasticity and peripheral nerve pathology are prominent neurological features, indicating a specific vulnerability of certain neuronal populations to autophagic disturbance. A typically biphasic disease course of late-onset neurodegeneration occurring on the background of a neurodevelopmental disorder further supports a role of autophagy in both neuronal development and maintenance. Additionally, an associated myopathy has been characterized in several conditions. The differential diagnosis comprises a wide range of other multisystem disorders, including mitochondrial, glycogen and lysosomal storage disorders, as well as ciliopathies, glycosylation and vesicular trafficking defects. The clinical overlap between the congenital disorders of autophagy and these conditions reflects the multiple roles of the proteins and/or emerging molecular connections between the pathways implicated and suggests an exciting area for future research. Therapy development for congenital disorders of autophagy is still in its infancy but may result in the identification of molecules that target autophagy more specifically than currently available compounds. The close connection with adult-onset neurodegenerative disorders highlights the relevance of research into rare early-onset neurodevelopmental conditions for much more common, age-related human diseases.Peer reviewe

Preclinical development of a vaccine against oligomeric alpha-synuclein based on virus-like particles

Parkinson's disease (PD) is a progressive and currently incurable neurological disorder characterised by the loss of midbrain dopaminergic neurons and the accumulation of aggregated alpha-synuclein (a-syn). Oligomeric a-syn is proposed to play a central role in spreading protein aggregation in the brain with associated cellular toxicity contributing to a progressive neurological decline. For this reason, a-syn oligomers have attracted interest as therapeutic targets for neurodegenerative conditions such as PD and other alpha-synucleinopathies. In addition to strategies using small molecules, neutralisation of the toxic oligomers by antibodies represents an attractive and highly specific strategy for reducing disease progression. Emerging active immunisation approaches using vaccines are already being trialled to induce such antibodies. Here we propose a novel vaccine based on the RNA bacteriophage (Qbeta) virus-like particle conjugated with short peptides of human a-syn. High titres of antibodies were successfully and safely generated in wild-type and human a-syn over-expressing (SNCA-OVX) transgenic mice following vaccination. Antibodies from vaccine candidates targeting the C-terminal regions of a-syn were able to recognise Lewy bodies, the hallmark aggregates in human PD brains. Furthermore, antibodies specifically targeted oligomeric and aggregated a-syn as they exhibited 100 times greater affinity for oligomeric species over monomer a-syn proteins in solution. In the SNCA-OVX transgenic mice used, vaccination was, however, unable to confer significant changes to oligomeric a-syn bioburden. Similarly, there was no discernible effect of vaccine treatment on behavioural phenotype as compared to control groups. Thus, antibodies specific for oligomeric a-syn induced by vaccination were unable to treat symptoms of PD in this particular mouse model.</p

The spectrum of neurodevelopmental, neuromuscular and neurodegenerative disorders due to defective autophagy

Primary dysfunction of autophagy due to Mendelian defects affecting core components of the autophagy machinery or closely related proteins have recently emerged as an important cause of genetic disease. This novel group of human disorders may present throughout life and comprises severe early-onset neurodevelopmental and more common adult-onset neurodegenerative disorders. Early-onset (or congenital) disorders of autophagy often share a recognizable "clinical signature," including variable combinations of neurological, neuromuscular and multisystem manifestations. Structural CNS abnormalities, cerebellar involvement, spasticity and peripheral nerve pathology are prominent neurological features, indicating a specific vulnerability of certain neuronal populations to autophagic disturbance. A typically biphasic disease course of late-onset neurodegeneration occurring on the background of a neurodevelopmental disorder further supports a role of autophagy in both neuronal development and maintenance. In addition, an associated myopathy has been characterized in several conditions. The differential diagnosis comprises a wide range of other multisystem disorders, including mitochondrial, glycogen and lysosomal storage disorders, as well as ciliopathies, glycosylation and vesicular trafficking defects. The clinical overlap between the congenital disorders of autophagy and these conditions reflects the multiple roles of the proteins and/or emerging molecular connections between the pathways implicated and suggests an exciting area for future research. Therapy development for congenital disorders of autophagy is still in its infancy but may result in the identification of molecules that target autophagy more specifically than currently available compounds. The close connection with adult-onset neurodegenerative disorders highlights the relevance of research into rare early-onset neurodevelopmental conditions for much more common, age-related human diseases

Future research directions on the "elusive" white shark

White sharks, Carcharodon carcharias, are often described as elusive, with little information available due to the logistical difficulties of studying large marine predators that make long-distance migrations across ocean basins. Increased understanding of aggregation patterns, combined with recent advances in technology have, however, facilitated a new breadth of studies revealing fresh insights into the biology and ecology of white sharks. Although we may no longer be able to refer to the white shark as a little-known, elusive species, there remain numerous key questions that warrant investigation and research focus. Although white sharks have separate populations, they seemingly share similar biological and ecological traits across their global distribution. Yet, white shark’s behavior and migratory patterns can widely differ, which makes formalizing similarities across its distribution challenging. Prioritization of research questions is important to maximize limited resources because white sharks are naturally low in abundance and play important regulatory roles in the ecosystem. Here, we consulted 43 white shark experts to identify these issues. The questions listed and developed here provide a global road map for future research on white sharks to advance progress toward key goals that are informed by the needs of the research community and resource managers

The changing career paths of PhDs and postdocs trained at EMBL

Individuals with PhDs and postdoctoral experience in the life sciences can pursue a variety of career paths. Many PhD students and postdocs aspire to a permanent research position at a university or research institute, but competition for such positions has increased. Here, we report a time-resolved analysis of the career paths of 2284 researchers who completed a PhD or a postdoc at the European Molecular Biology Laboratory (EMBL) between 1997 and 2020. The most prevalent career outcome was Academia: Principal Investigator (636/2284=27.8% of alumni), followed by Academia: Other (16.8%), Science-related Non-research (15.3%), Industry Research (14.5%), Academia: Postdoc (10.7%) and Non-science-related (4%); we were unable to determine the career path of the remaining 10.9% of alumni. While positions in Academia (Principal Investigator, Postdoc and Other) remained the most common destination for more recent alumni, entry into Science-related Non-research, Industry Research and Non-science-related positions has increased over time, and entry into Academia: Principal Investigator positions has decreased. Our analysis also reveals information on a number of factors – including publication records – that correlate with the career paths followed by researchers

Preclinical development of a vaccine against oligomeric alpha-synuclein based on virus-like particles

<div><p>Parkinson's disease (PD) is a progressive and currently incurable neurological disorder characterised by the loss of midbrain dopaminergic neurons and the accumulation of aggregated alpha-synuclein (a-syn). Oligomeric a-syn is proposed to play a central role in spreading protein aggregation in the brain with associated cellular toxicity contributing to a progressive neurological decline. For this reason, a-syn oligomers have attracted interest as therapeutic targets for neurodegenerative conditions such as PD and other alpha-synucleinopathies. In addition to strategies using small molecules, neutralisation of the toxic oligomers by antibodies represents an attractive and highly specific strategy for reducing disease progression. Emerging active immunisation approaches using vaccines are already being trialled to induce such antibodies. Here we propose a novel vaccine based on the RNA bacteriophage (Qbeta) virus-like particle conjugated with short peptides of human a-syn. High titres of antibodies were successfully and safely generated in wild-type and human a-syn over-expressing (<i>SNCA</i>-OVX) transgenic mice following vaccination. Antibodies from vaccine candidates targeting the C-terminal regions of a-syn were able to recognise Lewy bodies, the hallmark aggregates in human PD brains. Furthermore, antibodies specifically targeted oligomeric and aggregated a-syn as they exhibited 100 times greater affinity for oligomeric species over monomer a-syn proteins in solution. In the SNCA-OVX transgenic mice used, vaccination was, however, unable to confer significant changes to oligomeric a-syn bioburden. Similarly, there was no discernible effect of vaccine treatment on behavioural phenotype as compared to control groups. Thus, antibodies specific for oligomeric a-syn induced by vaccination were unable to treat symptoms of PD in this particular mouse model.</p></div

Effects of long-term immunisation with Qb-PD vaccines on a-syn aggregates.

<p>Male and female <i>SNCA</i>-OVX mice received 20 μg of Qb, Qb-PD1, Qb-PD3 or PBS every two weeks for a month, followed by monthly injections for 12 months (total duration of immunisation: for 13 months). (A-D) Immunofluorescence analysis was performed to detect a-syn aggregates with PLA in the substantia nigra, striatum, hippocampus and cerebellum. PLA puncta were quantified using ImageJ. (E) PLA puncta in 26–36 TH positive neurons were quantified and expressed as average mean of puncta per positive cell ± SEM (n = 3–4 mice per group), ANOVA followed by Dunn’s post hoc test. Scale bar 50 μm.</p

Assessment of behavioural effects of Qb-PD vaccines following long-term immunisation.

<p>Male and female <i>SNCA</i>-OVX mice received 20 μg of Qb, Qb-PD1, or Qb-PD3, or PBS every two weeks for a month, followed by monthly injections for 12 months (total duration of immunisation: 13 months). Parameters examined were (A) Rotarod fall latency, (B) spontaneous locomotor activity, (C) gait cadence, (D) forelimb stride length, (E) forelimb swing speed, (F) dry stool weight, (G) stool water content and (H) stool pellet number. Data are expressed as mean ± SEM (n = 13–16 mice per group).</p

Recognition of a-syn in human and mouse brain tissue.

<p>(A) Immunohistochemistry on paraffin-embedded brain tissue from a PD patient and control individual. Purified IgGs (1 mg/mL) used at 1:1000, commercial SYN211 antibody used at 1:2000. Region sampled, substantia nigra. Immunofluorescence on free-floating sections from the (B) striatum and (C) substantia nigra of 3 month old <i>SNCA</i>-OVX mice. Purified IgGs (1 mg/mL) used at 1:250, commercial BD a-syn antibody used at 1:500. Scale bars, 50 μm. (D) WB on 20 μg of striatum brain lysate from 3 month old mice. Samples from <i>Snca-/-</i> mice, mice over-expressing a-syn at moderate levels (line 21) and mice over-expressing a-syn with a two-fold increase (<i>SNCA</i>-OVX) were loaded (left-right). The MJFR1 antibody was used at 1:1000, while IgGs (1 mg/mL) from vaccinated mice were used at 1:500. Actin was used a loading control.</p

Competition ELISA to estimate relative affinity of vaccine-induced antibodies.

<p>Antigens in the form of peptides or recombinant monomer/oligomer protein preparations as described in the methods were coated on to microtitre plates and ELISA reading measured with fixed concentrations of serum IgG antibodies (at OD50 dilution) purified from pooled sera of vaccinated mice (n = 4), preincubated with serial dilutions of (A) free a-syn peptides (PD1, left; PD3, right); (B) free a-syn protein, either monomeric (left panels) or oligomeric (right panels) species, PD1 (top) and PD3 (bottom); and (C) liberated a-syn from haemolysed RBCs (orange line) or PBS negative control (blue line) for PD1 (top) and PD3 (bottom).</p