The SorLA protein, encoded by the SORL1 gene, is a major player in Alzheimer’s disease (AD) pathophysiology. Functional and genetic studies demonstrated that SorLA deficiency results in increased production of Aβ peptides, and thus a higher risk of AD. A large number of SORL1 missense variants have been identified in AD patients, but their functional consequences remain largely undefined. Here, we identified a new pathophysiological mechanism, by which rare SORL1 missense variants identified in AD patients result in altered maturation and trafficking of the SorLA protein. An initial screening, based on the overexpression of 70 SorLA variants in HEK293 cells, revealed that 15 of them (S114R, R332W, G543E, S564G, S577P, R654W, R729W, D806N, Y934C, D1535N, D1545E, P1654L, Y1816C, W1862C, P1914S) induced a maturation and trafficking-deficient phenotype. Three of these variants (R332W, S577P, and R654W) and two maturation-competent variants (S124R and N371T) were further studied in details in CRISPR/Cas9-modified hiPSCs. When expressed at endogenous levels, the R332W, S577P, and R654W SorLA variants also showed a maturation defective profile. We further demonstrated that these variants were largely retained in the endoplasmic reticulum, resulting in a reduction in the delivery of SorLA mature protein to the plasma membrane and to the endosomal system. Importantly, expression of the R332W and R654W variants in hiPSCs was associated with a clear increase of Aβ secretion, demonstrating a loss-of-function effect of these SorLA variants regarding this ultimate readout, and a direct link with AD pathophysiology. Furthermore, structural analysis of the impact of missense variants on SorLA protein suggested that impaired cellular trafficking of SorLA protein could be due to subtle variations of the protein 3D structure resulting from changes in the interatomic interactions.

Background: Alzheimer disease (AD) is a common complex disorder with a high genetic component. Loss-of-function (LoF)SORL1variants are one of the strongest AD genetic risk factors. Estimating their age-related penetrance is essential before putative use for genetic counseling or preventive trials. However, relative rarity and co-occurrence with the main AD risk factor,APOE-ε4, make such estimations difficult.Methods: We proposed to estimate the age-related penetrance ofSORL1-LoF variants through a survival framework by estimating the conditional instantaneous risk combining (i) a baseline for non-carriers ofSORL1-LoF variants, stratified byAPOE-ε4, derived from the Rotterdam study (N= 12,255), and (ii) an age-dependent proportional hazard effect forSORL1-LoF variants estimated from 27 extended pedigrees (including 307 relatives ≥ 40 years old, 45 of them having genotyping information) recruited from the French reference center for young Alzheimer patients. We embedded this model into an expectation-maximization algorithm to accommodate for missing genotypes. To correct for ascertainment bias, proband phenotypes were omitted. Then, we assessed if our penetrance curves were concordant with age distributions ofAPOE-ε4-stratifiedSORL1-LoF variant carriers detected among sequencing data of 13,007 cases and 10,182 controls from European and American case-control study consortia.Results: SORL1-LoF variants penetrance curves reached 100% (95% confidence interval [99–100%]) by age 70 amongAPOE-ε4ε4 carriers only, compared with 56% [40–72%] and 37% [26–51%] in ε4 heterozygous carriers and ε4 non-carriers, respectively. These estimates were fully consistent with observed age distributions ofSORL1-LoF variant carriers in case-control study data.Conclusions: We conclude thatSORL1-LoF variants should be interpreted in light ofAPOEgenotypes for future clinical applications.

Verletzungen des Ramus externus des N. laryngeus superior (RELS) können zum Verlust der Stimmhöhe sowie zu einer schnellen Ermüdbarkeit der Stimme führen. Dies kann insbesondere für Patienten, die von Berufs wegen auf ihre Stimmleistung angewiesen sind, dramatische Konsequenzen haben. Wir benutzten das Neuromonitoringgerät Neurosign 100® zur intraoperativen Identifikation des RELS. Bei 108 Patienten konnten wir 157 Nerven identifizieren, die Identifikationsrate lag bei 98,7 %. Wir konnten zeigen, dass die Ableitelektrode für das Neuromonitoring sowohl im M. cricothyreoideus als auch im M. vocalis positioniert werden kann. Die Nadellage im M. vocalis erleichtert das Neuromonitoring, wenn gleichzeitig auch ein Neuromonitoring des N. laryngeus recurrens durchgeführt wird. In 16 % unserer Fälle verlief der Nerv in unmittelbarer Nähe des oberen Schilddrüsenpols an oder zwischen den Ästen der A. thyreoidea superior und war deshalb hochgradig gefährdet, eine Läsion während der Versorgung der oberen Polgefäße zu erleiden. Unsere Daten bestätigen die Ergebnisse früherer und weniger umfangreicher Studien, bei denen ein solcher Nervenverlauf in 12 bis 14 % der Patienten beschrieben wird. Das Neuromonitoring erwies sich in unseren Händen als eine verlässliche und einfach durchzuführende Methode, um diesen Nerven zu identifizieren, und stellt unseres Erachtens damit ein wesentliches Element in Konzepten zur Schonung dieses Nerven dar. Injury to the external branch of the superior laryngeal nerve (EBSLN) during thyroid surgery can cause serious consequences for patients who depend on control of pitch and a clear and forceful voice, like singers or professional speakers. We used the Neurosign 100® nerve monitor to identify 157 nerves in 108 patients undergoing thyroid surgery. The EBSLN was successfully identified in 98.7 % of cases. The recording electrode could be placed either into the cricothyroid muscle or the vocal cord. The latter position proved superior if the recurrent laryngeal nerve had to be identified as well. 16 percent of the nerves crossed the branches of the superior thyroid artery at or below the upper pole of the gland, posing a “high risk” for intraoperative lesions. Our data confirm the results of smaller studies reporting this type of nerve course in 12 % to 14 % of patients. The present findings show a significant number of EBSLN to be in danger of injury when the superior thyroid artery is ligated during thyroid surgery. Neuromonitoring proofed to be a reliable method to identify the nerve, which is an important element in concepts to prevent its injury.

The SorLA protein, encoded by the SORL1 gene, is a major player in Alzheimer’s disease (AD) pathophysiology. Functional and genetic studies demonstrated that SorLA deficiency results in increased production of Aβ peptides, and thus a higher risk of AD. A large number of SORL1 missense variants have been identified in AD patients, but their functional consequences remain largely undefined. Here, we identified a new pathophysiological mechanism, by which rare SORL1 missense variants identified in AD patients result in altered maturation and trafficking of the SorLA protein. An initial screening, based on the overexpression of 71 SorLA variants in HEK293 cells, revealed that 15 of them (S114R, R332W, G543E, S564G, S577P, R654W, R729W, D806N, Y934C, D1535N, D1545E, P1654L, Y1816C, W1862C, P1914S) induced a maturation and trafficking-deficient phenotype. Three of these variations (R332W, S577P, and R654W) and two maturation-competent variations (S124R and N371T) were further studied in details in CRISPR/Cas9-modified hiPSCs. When expressed at endogenous levels, the R332W, S577P, and R654W SorLA variants also showed a maturation defective profile. We further demonstrated that these variants were largely retained in the endoplasmic reticulum, resulting in a reduction in the delivery of SorLA mature protein to the plasma membrane and to the endosomal system. Importantly, expression of the R332W and R654W variants in hiPSCs was associated with a clear increase of Aβ secretion, demonstrating a loss-of-function effect of these SorLA variants regarding this ultimate readout, and a direct link with AD pathophysiology. Furthermore, structural analysis of the impact of missense variations on SorLA protein indicated that impaired cellular trafficking of SorLA protein could be due to subtle variations of the protein 3D structure resulting from changes in the interatomic interactions.

La maladie de Huntington est neurodégénérative, génétique, rare, multifacette et de durée d'évolution longue, induisant une grande. Les biothérapies en cours d'essai sont réalisées sur des petits effectifs, avec un effet mesurable à long terme et hétérogène. Identifier des marqueurs d'évolution de la maladie et de réponse au traitement permettrait de mieux comprendre et d'améliorer les résultats des futurs essais cliniques. Nous avons développé une méthode de clustering pour l'efficacité d'un traitement dans le cadre de données longitudinales afin de définir des répondeurs et non répondeurs au traitement. Notre méthode, robuste pour les petits effectifs, combine un modèle linéaire mixte à deux pentes et un algorithme de clustering. Le modèle mixte génère des effets aléatoires, associés à la réponse au traitement, propres à chaque patient. L'algorithme de clustering permet de définir des sous-groupes selon la valeur des effets aléatoires. Trouver des sous-groupes de patients répondeurs permet de définir des marqueurs prédictifs de la réponse au traitement qui seront utilisés pour donner le traitement le mieux adapté à chaque patient. Nous avons discuté de l'intégration (i) des marqueurs prédictifs dans les plans expérimentaux des essais cliniques, en évaluant leur impact sur la puissance de l'étude; et (ii) des marqueurs pronostiques, en étudiant l¿impact du polymorphisme COMT sur le déclin cognitif des patients. Enfin, nous avons évalué l'effet d'apprentissage des tests neuropsychologiques, et montré comment une double évaluation à l'inclusion dans un essai clinique permettait de s'en affranchir quand le critère de jugement principal est le déclin cognitif.