Alzheimer's Disease Drug Development Research Articles

In vitro anticholinesterase potential of some spices consumed in Cameroon and their protective effects on hydrogen peroxide-mediated oxidative stress damage in SK-N-SH cells.

BackgroundMany neurodegenerative such as Alzheimer's disease (AD) are characterized by cholinergic dysfunction and oxidative stress which is a key event in neuronal death process. Thus, anticholinesterase and anti-oxidation compounds are two promising strategies in the development of AD drugs. Beyond their culinary use, spices are today studies for health purpose. In this study, some spices consumed in Cameroon were evaluated for their anticholinesterase and neuroprotective effects.MethodsColorimetric methods were used to determine total flavonoid and alkaloid content of a combinated extract (hydroethanolic + ethanolic extracts) of different selected spices. Aftermaths, anti-cholinesterase activity of spice extract was carried out using Ellman’s method. Finally, neuroprotective effects performed on human SK-N-SH cells stressed with H2O2 by assessing neuronal survival ( resazurin assay) and neuronal death (LDH assay).ResultsFlavonoid content of spices extract were ranged from 22.94 to 32.01 mg EQ/g DM and alkaloid content were ranged from 320 to 896 mg EQu/g DM. Among the spices studied, Xylopia parviflora presented the greatest acetylcholinesterase inhibition with an IC50 = 14 µg/mL. In Cell culture experiments, pre-incubation of SK-N-SH cell with the selected spices at different concentrations were improved neuronal survival and reduced the percentage of neuronal cells dead.ConclusionThe present results reveal that selected spices consumed in Cameroon have good anticholinesterase activity as well as neuroprotective effect on SK-N-SH which may provide new natural compounds that could help in the management of Alzheimer's disease.

Anti-fibrillization effects of sulfonamide derivatives on α-synuclein and hyperphosphorylated tau isoform 1N4R

In contrast to Aβ plaques, the spatiotemporal distribution of neurofibrillary tangles of hyperphosphorylated tau (p-tau) predicts cognitive impairment in Alzheimer's disease (AD), underscoring the key pathological role of p-tau and the utmost need to develop AD therapeutics centering upon the control of p-tau aggregation and cytotoxicity. Our drug discovery program is focused on compounds that prevent the aggregation and cytotoxicity of p-tau moieties of the tau isoform 1N4R due to its prevalence (1 N) and long-distance trans-synaptic propagation (4R). We prepared and tested twenty-four newly synthesized small molecules representing the urea (1, 2, 3), sulfonylurea (4), and sulfonamide (5–24) series and evaluated their anti-aggregation effects with biophysical methods (thioflavin T and S fluorescence assays, transmission electron microscopy) and intracellular inclusion cell-based assays. Pre-evaluation was performed on alpha-synuclein (α-syn) to identify molecules to be challenged with p-tau. The sulfonamide derivatives 18 and 20 exhibited an anti-fribrillization activity on α-syn and p-tau. Sulfonamide compounds 18 and 20 reduced inclusion formation in M17D neuroblastoma cells that express inclusion-prone αSynuclein3K::YFP. This project advances new concepts in targeting prone-to-aggregate proteins such as α-syn and p-tau, and provides a molecular scaffold for further optimization and pre-clinical studies focused on AD drug development.

Tau Beats Amyloid in Predicting Brain Atrophy in Alzheimer Disease: Implications for Prognosis and Clinical Trials.

Several biomarkers have emerged in the past few decades to quantify pathologic brain changes related to Alzheimer disease (AD). In particular, PET radiotracers that bind selectively to amyloid-β plaques and tau neurofibrillary tangles have advanced AD research and drug development by enabling the

A Phenotypic‐based Drug Discovery Approach to Inhibit the Misfolding of Hyperphosphorylated Tau Isoform 1N4R

The vast majority of Alzheimer’s disease (AD) drug development efforts have failed because of significant gap in the understanding of the pathophysiological processes and insufficient effort in identifying, optimizing, and validating small molecules inhibitors of aggregation. AD involves the misfolding of amyloid‐beta which forms Aβ plaques and, subsequently, activates different types of microglial cells leading to a pro‐inflammatory state. Microglia when activated have been known to increase tau phosphorylation and drive the trans‐synaptic spread of aggregated hyperphosphorylated tau (p‐tau). Most importantly, the spatiotemporal distribution of neurofibrillary tangles predicts cognitive impairment. Instead of the unmodified tau that shows minimal effects on cell viability, our research focuses on the control of p‐tau that possesses disease‐relevant molecular features, including hyperphosphorylation and cytotoxicity. Our drug discovery project is centered on the isoform 1N4R due to its prevalence (1N) and long‐distance trans‐synaptic propagation (4R). We identified compounds that inhibit the aggregation and cytotoxicity of p‐tau, which represent a novel approach for AD drug discovery and development. Due to lack of binding pocket(s) and fast‐switching conformation of p‐tau, we opted for a phenotypic‐based drug screening approach to identify and optimize hit candidates. In our current project, the anti‐aggregation activity and cellular protection of newly synthesized small molecules were evaluated with biophysical methods (thioflavin S fluorescence assays, photo‐induced cross‐linking of unmodified proteins, dynamic light scattering) and survival assays with human neuroblastoma SH‐SY5Y cells. We prepared and tested 3 different series of small molecules and challenged these molecules with our first‐tier screening assay, i.e. thioflavin S fluorescence assays. Ureas and sulfonylureas series failed to inhibit the aggregation of p‐tau 1N4R. The best prototypical representative of the sulfonamide series, compound JF‐19‐73, effectively antagonized the cytotoxicity of p‐tau in vitro at low micromolar concentrations, suggesting the potential for neuroprotection in vivo. Other representatives of the sulfonamide series exhibited an intermediate effect to antagonize the aggregation and cytotoxicity of p‐tau isoform 1N4R. This project opens doors in targeting undruggable prone‐to‐aggregate proteins such as p‐tau and provides a molecular scaffold for further optimization and pre‐clinical studies focused on AD drug development.

Altered protein expression of membrane transporters in isolated cerebral microvessels and brain cortex of a rat Alzheimer's disease model

There is growing evidence that membrane transporters expressed at the blood-brain barrier (BBB) and brain parenchymal cells play an important role in Alzheimer's disease (AD) development and progression. However, quantitative information about changes in transporter protein expression at neurovascular unit cells in AD is limited. Here, we studied the changes in the absolute protein expression of five ATP-binding cassette (ABC) and thirteen solute carrier (SLC) transporters in the isolated brain microvessels and brain cortical tissue of TgF344-AD rats compared to age-matched wild-type (WT) animals using liquid chromatography tandem mass spectrometry based quantitative targeted absolute proteomic analysis. Moreover, sex-specific alterations in transporter expression in the brain cortical tissue of this model were examined. Protein expressions of Abcg2, Abcc1 and FATP1 (encoded by Slc27a1) in the isolated brain microvessels of TgF344-AD rats were 3.1-, 2.0-, 4.3-fold higher compared to WT controls, respectively (p < 0.05). Abcc1 and 4F2hc (encoded by Slc3a2) protein expression was significantly up-regulated in the brain cortical tissue of male TgF344-AD rats compared to male WT rats (p < 0.05). The study provides novel information for the elucidation of molecular mechanisms underlying AD and valuable knowledge about the optimal use of the TgF344-AD rat model in AD drug development and drug delivery research.

The synapse as a treatment avenue for Alzheimer’s Disease

Alzheimer's disease (AD) is a neurodegenerative disorder with devastating symptoms, including memory impairments and cognitive deficits. Hallmarks of AD pathology are amyloid-beta (Aβ) deposition forming neuritic plaques and neurofibrillary tangles (NFTs). For many years, AD drug development has mainly focused on directly targeting the Aβ aggregation or the formation of tau tangles, but this disease has no cure so far. Other common characteristics of AD are synaptic abnormalities and dysfunctions such as synaptic damage, synaptic loss, and structural changes in the synapse. Those anomalies happen in the early stages of the disease before behavioural symptoms have occurred. Therefore, better understanding the mechanisms underlying the synaptic dysfunction found in AD and targeting the synapse, especially using early treatment windows, can lead to finding novel and more effective treatments that could improve the lives of AD patients. Researchers have recently started developing different disease-modifying treatments targeting the synapse to rescue and prevent synaptic dysfunction in AD. The main objectives of these new strategies are to halt synaptic loss, strengthen synaptic connections, and improve synaptic density, potentially leading to the rescue or prevention of cognitive impairments. This article aims to address the mechanisms of synaptic degeneration in AD and discuss current strategies that focus on the synapse for AD therapy. Alzheimer's disease (AD) is a neurodegenerative disorder that significantly impairs memory and causes cognitive and behavioural deficits. Scientists worldwide have tried to find a treatment that can reverse or rescue AD symptoms, but there is no cure so far. One prominent characteristic of AD is the brain atrophy caused by significant synaptic loss and overall neuronal damage, which starts at the early stages of the disease before other AD hallmarks such as neuritic plaques and NFTs. The present review addresses the underlying mechanisms behind synaptic loss and dysfunction in AD and discusses potential strategies that target the synapse.

Alzheimer-like tau accumulation in dentate gyrus mossy cells induces spatial cognitive deficits by disrupting multiple memory-related signaling and inhibiting local neural circuit.

Abnormal tau accumulation and spatial memory loss constitute characteristic pathology and symptoms of Alzheimer disease (AD). Yet, the intrinsic connections and the mechanism between them are not fully understood. In the current study, we observed a prominent accumulation of the AD‐like hyperphosphorylated and truncated tau (hTau N368) proteins in hippocampal dentate gyrus (DG) mossy cells of 3xTg‐AD mice. Further investigation demonstrated that the ventral DG (vDG) mossy cell‐specific overexpressing hTau for 3 months induced spatial cognitive deficits, while expressing hTau N368 for only 1 month caused remarkable spatial cognitive impairment with more prominent tau pathologies. By in vivo electrophysiological and optic fiber recording, we observed that the vDG mossy cell‐specific overexpression of hTau N368 disrupted theta oscillations with local neural network inactivation in the dorsal DG subset, suggesting impairment of the ventral to dorsal neural circuit. The mossy cell‐specific transcriptomic data revealed that multiple AD‐associated signaling pathways were disrupted by hTau N368, including reduction of synapse‐associated proteins, inhibition of AKT and activation of glycogen synthase kinase‐3β. Importantly, chemogenetic activating mossy cells efficiently attenuated the hTau N368‐induced spatial cognitive deficits. Together, our findings indicate that the mossy cell pathological tau accumulation could induce the AD‐like spatial memory deficit by inhibiting the local neural network activity, which not only reveals new pathogenesis underlying the mossy cell‐related spatial memory loss but also provides a mouse model of Mossy cell‐specific hTau accumulation for drug development in AD and the related tauopathies.

Iron Dyshomeostasis and Ferroptosis: A New Alzheimer's Disease Hypothesis?

Iron plays a crucial role in many physiological processes of the human body, but iron is continuously deposited in the brain as we age. Early studies found iron overload is directly proportional to cognitive decline in Alzheimer’s disease (AD). Amyloid precursor protein (APP) and tau protein, both of which are related to the AD pathogenesis, are associated with brain iron metabolism. A variety of iron metabolism-related proteins have been found to be abnormally expressed in the brains of AD patients and mouse models, resulting in iron deposition and promoting AD progression. Amyloid β (Aβ) and hyperphosphorylated tau, two pathological hallmarks of AD, can also promote iron deposition in the brain, forming a vicious cycle of AD development-iron deposition. Iron deposition and the subsequent ferroptosis has been found to be a potential mechanism underlying neuronal loss in many neurodegenerative diseases. Iron chelators, antioxidants and hepcidin were found useful for treating AD, which represents an important direction for AD treatment research and drug development in the future. The review explored the deep connection between iron dysregulation and AD pathogenesis, discussed the potential of new hypothesis related to iron dyshomeostasis and ferroptosis, and summarized the therapeutics capable of targeting iron, with the expectation to draw more attention of iron dysregulation and corresponding drug development.

Structural Modifications on Chalcone Framework for Developing New Class of Cholinesterase Inhibitors.

Due to the multifaceted pharmacological activities of chalcones, these scaffolds have been considered one of the most privileged frameworks in the drug discovery process. Structurally, chalcones are α, β-unsaturated carbonyl functionalities with two aryl or heteroaryl units. Amongst the numerous pharmacological activities explored for chalcone derivatives, the development of novel chalcone analogs for the treatment of Alzheimer’s disease (AD) is among the research topics of most interest. Chalcones possess numerous advantages, such as smaller molecular size, opportunities for further structural modification thereby altering the physicochemical properties, cost-effectiveness, and convenient synthetic methodology. The present review highlights the recent evidence of chalcones as a privileged structure in AD drug development processes. Different classes of chalcone-derived analogs are summarized for the easy understanding of the previously reported analogs as well as the importance of certain functionalities in exhibiting cholinesterase inhibition. In this way, this review will shed light on the medicinal chemistry fraternity for the design and development of novel promising chalcone candidates for the treatment of AD.

Preliminary studies of an imidazole-based alcohol derivative for imaging of Heme oxygenase 1

Heme oxygenase-1 (HO-1) has been involved in the pathogenesis of Alzheimer's disease (AD), thus constituting a promising target for AD drug development. Positron emission tomography (PET) is a fully translational imaging technology, which will help us understand the role of HO-1 in the progression of AD, facilitating to validate promising HO-1 inhibitors in clinical trials. To our knowledge, there is no report on PET imaging probe targeting HO-1 in animals and humans. We report herein the synthesis and characterization of a 11C-labeled imidazole-based alcohol derivative ([11C]QC-33) for imaging of HO-1 in the brain. The desired product [11C]QC-33 was afforded with a radiochemical yield of 16 ± 9% (n = 3, decay corrected). The radiochemical purity was greater than 99%, and the molar radioactivity was greater than 185 GBq/μmol. In vitro autoradiography studies indicated specific binding of [11C]QC-33 in the HO-1 rich regions, showing 75%, 75%, and 69% radioactivity binding reductions in cerebellum, brain stem, and midbrain, respectively. PET/CT scanning in C57BL/6 mice showed low brain uptake and poor blood–brain barrier (BBB) penetration of [11C]QC-33. These results suggested that [11C]QC-33 can serve as a lead compound to advance the development of next generation PET tracer with the potential to monitor HO-1 in AD progression.

USP25 inhibition ameliorates Alzheimer's pathology through the regulation of APP processing and Aβ generation.

Down syndrome (DS), or trisomy 21, is one of the critical risk factors for early-onset Alzheimer’s disease (AD), implicating key roles for chromosome 21–encoded genes in the pathogenesis of AD. We previously identified a role for the deubiquitinase USP25, encoded on chromosome 21, in regulating microglial homeostasis in the AD brain; however, whether USP25 affects amyloid pathology remains unknown. Here, by crossing 5×FAD AD and Dp16 DS mice, we observed that trisomy 21 exacerbated amyloid pathology in the 5×FAD brain. Moreover, bacterial artificial chromosome (BAC) transgene–mediated USP25 overexpression increased amyloid deposition in the 5×FAD mouse brain, whereas genetic deletion of Usp25 reduced amyloid deposition. Furthermore, our results demonstrate that USP25 promoted β cleavage of APP and Aβ generation by reducing the ubiquitination and lysosomal degradation of both APP and BACE1. Importantly, pharmacological inhibition of USP25 ameliorated amyloid pathology in the 5×FAD mouse brain. In summary, we identified the DS-related gene USP25 as a critical regulator of AD pathology, and our data suggest that USP25 serves as a potential pharmacological target for AD drug development.

Delivery of Intravenously Administered Antibodies Targeting Alzheimer's Disease-Relevant Tau Species into the Brain Based on Receptor-Mediated Transcytosis.

Alzheimer’s disease (AD) is a neurodegenerative disease that causes memory loss, cognitive decline, and eventually dementia. The etiology of AD and its pathological mechanisms remain unclear due to its complex pathobiology. At the same time, the number of patients with AD is increasing worldwide. However, no therapeutic agents for AD are currently available for definitive care. Several phase 3 clinical trials using agents targeting amyloid β (Aβ) and its related molecules have failed, with the exception of aducanumab, an anti-Aβ monoclonal antibody (mAb), clinically approved by the US Food and Drug Administration in 2021, which could be modified for AD drug development due to controversial approval. Neurofibrillary tangles (NFTs) composed of tau rather than senile plaques composed of Aβ are correlated with AD pathogenesis. Moreover, Aβ and tau pathologies initially proceed independently. At a certain point in the progression of AD symptoms, the Aβ pathology is involved in the alteration and spreading of the tau pathology. Therefore, tau-targeting therapies have attracted the attention of pharmaceutical scientists, as well as Aβ-targeting therapies. In this review, I introduce the implementations and potential of AD immunotherapy using intravenously administered anti-tau and anti-receptor bispecific mAbs. These cross the blood-brain barrier (BBB) based on receptor-mediated transcytosis and are subsequently cleared by microglia based on Fc-mediated endocytosis after binding to tau and lysosomal degradation.

Multi-Targets: An Unconventional Drug Development Strategy for Alzheimer's Disease.

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder that eventually leads to dementia and death of the patient. Despite the enormous amounts of resources and efforts for AD drug development during the last three decades, no effective treatments have been developed that can slow or halt the progression of the disease. Currently available drugs for treating AD can only improve clinical symptoms temporarily with moderate efficacies. In recent years, the scientific community has realized these challenges and reconsidered the future directions of AD drug development. The most significant recent changes in AD drug development strategy include shifting from amyloid-based targets to other targets, such as tau, and efforts toward better designs for clinical trials. However, most AD drug development is still focused on a single mechanism or target, which is the conventional strategy for drug development. Although multifactorial mechanisms and, on this basis, multi-target strategies have been proposed in recent years, this approach has not been widely recognized and accepted by the mainstream of AD drug development. Here, we emphasize the multifactorial mechanisms of AD and discuss the urgent need for a paradigm shift in AD drug development from a single target to multiple targets, either with the multi-target–directed ligands approach or the combination therapy approach. We hope this article will increase the recognition of the multifactorial nature of AD and promote this paradigm shift. We believe that such a shift will facilitate successful development of effective AD therapies.

Structure of unmodified and pyroglutamylated amyloid beta peptides in lipid membranes

Alzheimer’s disease (AD) is a neurodegenerative disorder that causes brain atrophy, neuronal and synaptic loss, cognitive decline, and death. Worldwide, over 30 million people suffer from AD. Despite extensive basic and clinical research and multibillion investments in pharmaceutical efforts, no disease-modifying drugs are available for AD. One of the roadblocks in AD drug development is the incomplete understanding of molecular mechanisms underlying the disease. A major histopathological trait of AD is extracellular deposition of amyloid-beta (Aβ) peptide in brain parenchyma and vasculature, but the most toxic species are the intracellular oligomeric forms of Aβ.

Alzheimer's disease drug development pipeline: 2022.

IntroductionAlzheimer's disease (AD) represents a global health crisis. Treatments are needed to prevent, delay the onset, slow the progression, improve cognition, and reduce behavioral disturbances of AD. We review the current clinical trials and drugs in development for the treatment of AD.MethodsWe searched the governmental website clinicaltrials.gov where are all clinical trials conducted in the United States must be registered. We used artificial intelligence (AI) and machine learning (ML) approaches to ensure comprehensive detection and characterization of trials and drugs in development. We use the Common Alzheimer's Disease Research Ontology (CADRO) to classify drug targets and mechanisms of action of drugs in the pipeline.ResultsAs of January 25, 2022 (index date for this study) there were 143 agents in 172 clinical trials for AD. The pipeline included 31 agents in 47 trials in Phase 3, 82 agents in 94 trials in Phase 2, and 30 agents in 31 trials in Phase 1. Disease‐modifying therapies represent 83.2% of the total number of agents in trials; symptomatic cognitive enhancing treatments represent 9.8% of agents in trials; and drugs for the treatment of neuropsychiatric symptoms comprise 6.9%. There is a diverse array of drug targets represented by agents in trials including nearly all CADRO categories. Thirty‐seven percent of the candidate agents in the pipeline are repurposed drugs approved for other indications. A total of 50,575 participants are needed to fulfill recruitment requirements for all currently active clinical trials.DiscussionThe AD drug development pipeline has agents representing a substantial array of treatment mechanisms and targets. Advances in drug design, outcome measures, use of biomarkers, and trial conduct promise to accelerate the delivery of new and better treatments for patients with AD.Highlights There are 143 drugs in the current Alzheimer's disease (AD) drug development pipeline.Disease‐modifying therapies represent 83.2% of the candidate treatments.Current trials require 50,575 participants who will donate 3,878,843 participant‐weeks to clinical trials.The biopharmaceutical industry sponsors 50% of all clinical trials including 68% of Phase 3 trials.Sixty‐three percent of Phase 3 trials and 46% of Phase 2 trials include non–North American clinical trial site locations indicating the global ecosystem required for AD drug development.

In Situ Imaging of Formaldehyde in Live Mice with High Spatiotemporal Resolution Reveals Aldehyde Dehydrogenase-2 as a Potential Target for Alzheimer's Disease Treatment.

Alterations in formaldehyde (FA) homeostasis are associated with the pathology of Alzheimer's disease (AD). In vivo tracking of FA flux is important for understanding the underlying molecular mechanisms, but is challenging due to the lack of sensitive probes favoring a selective, rapid, and reversible response toward FA. In this study, we re-engineered the promiscuous and irreversible phenylhydrazines to make them selective and reversible toward FA by tuning their nucleophilicity. This effort resulted in PFM309, a selective (selectivity coefficient KFA,methylglyoxal = 0.06), rapid (t1/2 = 32 s at [FA] = 200 μM), and reversible fluorogenic probe (K = 6.24 mM-1) that tracks the FA flux in both live cells and live mice. In vivo tracking of the FA flux was realized by PFM309 imaging, which revealed the gradual accumulation of FA in the live mice brain during normal aging and its further increase in AD mice. We further identified the age-dependent loss of catabolism enzymes ALDH2 and ADH5 as the primary mechanism responsible for formaldehyde excess. Activating ALDH2 with the small molecular activator Alda1 significantly protected neurovascular cells from formaldehyde overload and consequently from impairment during AD progress both in vitro and in vivo. These findings revealed PFM309 as a robust tool to study AD pathology and highlight ALDH2 as a potential target for AD drug development.

Identification of a Thyroid Hormone Derivative as a Pleiotropic Agent for the Treatment of Alzheimer's Disease.

The identification of effective pharmacological tools for Alzheimer’s disease (AD) represents one of the main challenges for therapeutic discovery. Due to the variety of pathological processes associated with AD, a promising route for pharmacological intervention involves the development of new chemical entities that can restore cellular homeostasis. To investigate this strategy, we designed and synthetized SG2, a compound related to the thyroid hormone thyroxine, that shares a pleiotropic activity with its endogenous parent compound, including autophagic flux promotion, neuroprotection, and metabolic reprogramming. We demonstrate herein that SG2 acts in a pleiotropic manner to induce recovery in a C. elegans model of AD based on the overexpression of Aβ42 and improves learning abilities in the 5XFAD mouse model of AD. Further, in vitro ADME-Tox profiling and toxicological studies in zebrafish confirmed the low toxicity of this compound, which represents a chemical starting point for AD drug development.

The Critical Path for Alzheimer’s Disease (CPAD) Consortium: A platform for pre‐competitive data sharing, standardization, and analysis to support quantitative tools for AD drug development

AbstractBackgroundThe primary objective of the Critical Path for Alzheimer’s Disease (CPAD) consortium, a pre‐competitive consortium of the Critical Path Institute, is to promote, support, and manage pre‐competitive data and knowledge sharing from Alzheimer disease (AD) drug development stakeholders. Since 2019, CPAD has embarked on a new data sharing initiative to acquire fluid and imaging biomarker‐rich data sources for development of novel tools to quantify Alzheimer’s Disease progression across the AD continuum.MethodPatient‐level data from contemporary Phase2/Phase3 AD clinical trials and observational studies is acquired through formal data contribution agreements within CPAD, curated in collaboration with data contributors, standardized to common structure and terminology, and input into a relational data model (RDM) tool to generate analysis subsets. Data standardization is being accomplished using Clinical Data Interchange Standards Consortium (CDISC) standards. The RDM was developed using the open‐source software R and converts CDISC structure to analysis ready files for quantitative analyses through user defined primary keys, i.e. unique identifying variables for extracting information. The RDM includes a front‐end data interrogator that provides users with versatile capabilities to visualize and tabulate the data.ResultAs of October 2020, CPAD’s database (https://codr.c‐path.org/) contains 41 studies, standardized to the AD CDISC standards, of which three datasets were used to generate analysis subsets. Four additional datasets were recently acquired and are being standardized. Execution of Data Contribution Agreements (DCAs) is in progress for two contemporary datasets rich in key fluid and imaging biomarkers. Nine additional prioritized datasets are targeted for acquisition. The RDM is currently being used to transform the CDISC standardized data into analysis ready subsets containing patient demographics (age, sex), fluid biomarker information (Aβ 40/42), and clinical outcome assessments (Clinical Dementia Rating Scale – Sum of Boxes).ConclusionThe data acquisition and analysis platform developed is stimulating pre‐competitive efforts to generate quantitative tools for advancing AD drug development. Previously developed tools, including a regulatory endorsed clinical trial simulation tool, help optimize trial design and advance drug development in AD.

Single cell transcriptomic profiling of tau pathophysiology in a novel 3D neural-glial coculture model.

Research into neurodegeneration has been hampered by a lack of systems that rapidly and accurately recapitulate neurodegenerative processes. We report development of AstAD, a model combining human iPSC derived neurons (hiNCs) and astrocytes (hiACs) in self-aggregating 3D organoids that reproducibly develop a robust range of tau pathology and neurodegeneration. AstAD is a scalable and highly manipulatable model of neurodegeneration and critical understudied glial responses. Additionally, the AstAD model induces pathology without reliance on genetic overexpression. AstAD represents an important technological advance in modeling Alzheimer's Disease (AD), and is a promising tool for identifying disease modifying treatments. The AstAD system was developed by combining advances in 3D cell modeling and tau propagation. We perform directed differentiation of wild-type iPSC to hiNCs and hiACs, and hiNCs are selectively exposed to toxic aggregations to induce pathology. Cells are combined in microwells to induce self-aggregation, and pathology develops over the course of three weeks. Tau pathogenesis in the AstAD model was correlated with human tauopathy progression by immunopathology and single cell transcriptomics, and the application of a selective inhibitor PU-H71 demonstrated the efficacy of the model to explore disease modifying treatment. The AstAD model consistently displays a robust range of tau pathology within three weeks, including progressive development of hyperphosphorylated tau at positions T181, S262, and S396/404, misfolded tau (MC1, TOMA2), and fibrillar tau (THIOS). Neurodegeneration (FlouroJadeB, LDH) is accompanied by reactive astrogliosis (GFAP), which single cell transcriptomics reveals to be driven by heat shock proteins (HSPs). The observed tau pathology and transcriptional responses were reduced by treatment with PU-H71, which eliminates dysfunctional HSP epichaperome. Together this evidence demonstrates a rapid and reversible recapitulation of tau pathogenesis. The novel AstAD model consistently recapitulates tau-induced neurodegeneration in a human system, and allows for the elucidation of neuronal and astrocytic responses. Additionally, we have demonstrated the power of AstAD to model the reduction of tau pathology by drug treatment. Continued expansion of the AstAD model will include integration of microglia. The AstAD model represents a technological advance that will allow for the rapid recapitulation of neurodegeneration in a manipulatable system that promises to accelerate drug development for AD.

Human serum albumin fusion protein as therapeutics for targeting amyloid beta in Alzheimer’s diseases

Alzheimer's disease (AD) is characterized by amyloid beta (Aβ) plaques and neurofibrillary tangles. AD drug development has been limited due to the presence of the blood–brain barrier (BBB), which prevents efficient uptake of therapeutics into the brain. To solve this problem, we used trans-activator of transcription (TAT)-transducing domain and added the human serum albumin (HSA) carrier to increase the half-life of the drug within the body. In addition, we included the protein of interest for lowering Aβ deposition and/or neurofibrillary tangles. We made HSA fusion protein (designated AL04) which contains Cystatin C (CysC) as core mechanism of action moiety in the construct containing tandem repeat TAT (dTAT). After purification of 80KDa AL04, we investigate the therapeutic potential of AL04 in vitro and AD mouse model Tg2576. We evaluated the permeability of AL04 through the BBB using a cell-basedhuman BBB model and show that dTAT plays a role in facilitating the delivery of 80 kDa protein. We found out that AL04 attenuates Aβ-induced neurotoxicity in PC12 cells. In Tg2576 mice brain, Aβ plaques were dramatically reduced in AL04 treated mice. These data suggest that BBB-crossing albumin fusion protein AL04 with CysC active moiety can be a disease modifying treatment for AD.