Kondru, Naveen

AbstractBackgroundDementia with Lewy bodies (DLB), the second most prevalent form of degenerative dementia, is characterized by pathological α‐synuclein (αSyn) protein aggregation in neurons. Pathological αSyn shows a protein‐seeding phenomenon that may result in aggregation years before clinical symptoms. Studies using Real‐time quaking‐induced conversion (RT‐QuIC) assay show αSyn seeding from peripheral tissue including skin and cerebrospinal Fluid (CSF). Identifying early biomarkers for DLB is essential for accurate diagnosis, and developing effective treatments to slow progression. Herein, we investigate αSyn seeding activities in CSF from patients with mild cognitive impairment with Lewy bodies (MCI‐LB) phenoconverting to DLB.MethodCSF samples were obtained from 3 MCI‐LB and 2 DLB patients at baseline and two‐year follow‐up appointments. CSF samples were tested for their potential to seed α‐synuclein aggregation using an RT‐QuIC assay. CSF samples were tested by seeding 2 μl of CSF in 98 ul of RT‐QuIC reaction mixture with human recombinant αSyn protein, analyzed on a FLUOstar Omega plate reader. Results were recorded every 30 minutes over a period of 45 hours analyzed based on the reaction kinetics to determine the seeding ability of the α‐synuclein in the CSF samples.ResultWe observed seed amplification at first visit for both DLB patients and one MCI‐LB patient. The absolute quantification of aggregation increased at second visit (2 years apart) for one DLB patient and one MCI‐LB. The other two MCI‐LB patients showed no amplification at baseline visit but showed signal at two‐year visit, by which time all three had phenoconverted to DLB. Interestingly the second DLB patient showed a decline at two‐year follow‐up. The medical and neuroimaging records of that patient indicate that the patient’s cognitive decline began as early‐onset disease (first symptoms at age 50 years).ConclusionWe demonstrate that measures of seed aggregation in CSF with RT‐QuIC are informative in MCI‐LB and DLB, and may also reflect changes in disease progression and dynamics.

Pick’s disease (PiD) is a devastating neurodegenerative disease that is characterized by dementia, frontotemporal lobar degeneration, and the aggregation of 3R tau in pathognomonic inclusions known as Pick bodies. The term PiD has adopted many meanings since its conception in 1926, but it is currently used as a strictly neuropathological term, since PiD patients cannot be diagnosed during life. Due to its rarity, PiD remains significantly understudied, and subsequently, the etiology and pathomechanisms of the disease remain to be elucidated. The study of PiD and the preferential 3R tau accumulation that is unique to PiD is imperative in order to expand the current understanding of the disease and inform future studies and therapeutic development, since the lack of intervention strategies for tauopathies remains an unmet need. Yet, the lack of an antemortem diagnostic test for the disease has further complicated the study of PiD. The development of a clinical diagnostic assay for PiD will be a vital step in the study of the disease that will greatly contribute to therapeutic research, clinical trial design and patient recruitment and ultimately improve patient outcomes. Seed aggregation assays have shown great promise for becoming ante mortem clinical diagnostic tools for many proteinopathies, including tauopathies. Future research on adapting and optimizing current seed aggregation assays to successfully detect 3R tau pathogenic forms from PiD samples will be critical in establishing a 3R tau specific seed aggregation assay that can be used for clinical diagnosis and treatment evaluation.

Abstract Synucleinopathies are clinically and pathologically heterogeneous disorders characterized by pathologic aggregates of α-synuclein in neurons and glia, in the form of Lewy bodies, Lewy neurites, neuronal cytoplasmic inclusions, and glial cytoplasmic inclusions (GCIs). Synucleinopathies can be divided into two major disease entities: Lewy body disease (LBD) and multiple system atrophy (MSA). Common clinical presentations of LBD are Parkinson's disease (PD), PD with dementia (PDD), and dementia with Lewy bodies (DLB), while MSA has two major clinical subtypes, MSA with predominant cerebellar ataxia (MSA-C) and MSA with predominant parkinsonism (MSA-P). There are currently no disease-modifying therapies for the synucleinopathies, but elucidation of genetics and mechanisms of α-synuclein conversion to pathologic oligomers and insoluble fibrils offer hope for eventual therapies. It remains unclear how α-synuclein can be associated with distinct cellular pathologies (e.g., Lewy bodies and GCI) and what factors determine neuroanatomical and cell type vulnerability. Accumulating evidence from in vitro and in vivo experiments suggests that α-synuclein species derived from LBD and MSA are distinct "strains" having different seeding properties. Recent advancements in in vitro seeding assays, such as real-time quaking-induced conversion (RT-QuIC) and protein misfolding cyclic amplification (PMCA), not only demonstrate distinct seeding activity in the synucleinopathies, but also offer exciting opportunities for molecular diagnosis using readily accessible peripheral tissues. Cryogenic electron microscopy (cryo-EM) structural studies of α-synuclein derived from recombinant or brain-derived filaments provide new insight into mechanisms of seeding in synucleinopathies. In this review, we describe clinical, genetic and neuropathologic features of synucleinopathies, including a review of classification and staging schemes for LBD. We also review evidence supporting the existence of distinct α-synuclein strains in LBD and MSA.

ABSTRACT Background An unmet clinical need in Parkinson's disease (PD) is to identify biomarkers for diagnosis, preferably in peripherally accessible tissues such as skin. Immunohistochemical studies have detected pathological α‐synuclein (αSyn) in skin biopsies from PD patients albeit sensitivity needs to be improved. Objective Our study provides the ultrasensitive detection of pathological αSyn present in the skin of PD patients, and thus, pathological αSyn in skin could be a potential biomarker for PD. Methods The real‐time quaking‐induced conversion assay was used to detect pathological αSyn present in human skin tissues. Further, we optimized this ultra‐sensitive and specific assay for both frozen and formalin‐fixed paraffin‐embedded sections of skin tissues. We determined the seeding kinetics of the αSyn present in the skin from autopsied subjects consisting of frozen skin tissues from 25 PD and 25 controls and formalin‐fixed paraffin‐embedded skin sections from 12 PD and 12 controls. Results In a blinded study of skin tissues from autopsied subjects, we correctly identified 24/25 PD and 24/25 controls using frozen skin tissues (96% sensitivity and 96% specificity) compared to 9/12 PD and 10/12 controls using formalin‐fixed paraffin‐embedded skin sections (75% sensitivity and 83% specificity). Conclusions Our blinded study results clearly demonstrate the feasibility of using skin tissues for clinical diagnosis of PD by detecting pathological αSyn. Moreover, this peripheral biomarker discovery study may have broader translational value in detecting misfolded proteins in skin samples as a longitudinal progression marker. © 2020 International Parkinson and Movement Disorder Society