Cathryn Haigh

National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States · Rocky Mountain Laboratories

Prion diseases are transmissible, fatal neurologic diseases of mammals caused by the accumulation of mis-folded, disease associated prion protein (PrPd). Creutzfeldt-Jakob Disease (CJD) is the most common human prion disease and can occur by sporadic onset (sCJD) (~85% of CJD cases), genetic mutations in the prion protein gene (10–15%) or iatrogeni...

Proteolytic cell surface release (‘shedding’) of the prion protein (PrP), a broadly expressed GPI-anchored glycoprotein, by the metalloprotease ADAM10 impacts on neurodegenerative and other diseases in animal and in vitro models. Recent studies employing the latter also suggest shed PrP (sPrP) to be a ligand in intercellular communication and criti...

Chronic wasting disease (CWD) is a cervid prion disease with unknown zoonotic potential that might pose a risk to humans who are exposed. To assess the potential of CWD to infect human neural tissue, we used human cerebral organoids with 2 different prion genotypes, 1 of which has previously been associated with susceptibility to zoonotic prion dis...

Background Sporadic Creutzfeldt-Jakob disease (sCJD), the most common human prion disease, is a fatal neurodegenerative disease with currently no treatment options. Stem cell therapy for neurodegenerative diseases is emerging as a possible treatment option. However, while there are a few clinical trials for other neurodegenerative disorders such as...

Proteolytic cell surface release (‘shedding’) of the prion protein (PrP), a broadly expressed GPI-anchored glycoprotein, by the metalloprotease ADAM10 impacts on neurodegenerative and other diseases in animal and in vitro models. Recent studies employing the latter also suggest shed PrP (sPrP) to be a ligand in intercellular communication and criti...

Mis-folding of the prion protein (PrP) is known to cause neurodegenerative disease, however the native function of this protein remains poorly defined. PrP has been linked with many cellular functions, including cellular proliferation and senescence. It is also known to influence epidermal growth factor receptor (EGFR) signalling, a pathway that is...

Abnormal deposition of α-synuclein is a key feature and biomarker of Parkinson’s disease. α-Synuclein aggregates can propagate themselves by a prion-like seeding-based mechanism within and between tissues and are hypothesized to move between the intestine and brain. α-Synuclein RT-QuIC seed amplification assays have detected Parkinson’s-associated...

Cellular prion protein (PrPC) protects neurons against oxidative stress damage. This role is lost upon its misfolding into insoluble prions in prion diseases, and correlated with cytoskeletal breakdown and neurophysiological deficits. Here we used mouse neuronal models to assess how PrPC protects the neuronal cytoskeleton, and its role in network c...

Human cerebral organoids (COs) are three-dimensional self-organizing cultures of cerebral brain tissue differentiated from induced pluripotent stem cells. We have recently shown that COs are susceptible to infection with different subtypes of Creutzfeldt–Jakob disease (CJD) prions, which in humans cause different manifestations of the disease. The...

Fatal familial insomnia (FFI) is a rare neurodegenerative disease caused by a dominantly inherited single amino acid substitution (D178N) within the prion protein (PrP). No in vitro human brain tissue model for this disease has previously been available. Consequently, how this mutation exerts its damaging effect on brain cells is still unknown. Usi...

Stem cells have the capacity to differentiate into the mature cells of any organ within the body. For this reason, they offer an interesting opportunity to model many cellular systems and their associated diseases. Prion diseases (PrDs) are a family of fatal neurodegenerative diseases caused by mis-folding of the prion protein (PrP), a protein lack...

Prion diseases are a group of rare, transmissible, and invariably fatal neurodegenerative diseases that affect both humans and animals. The cause of these diseases is misfolding of the prion protein into pathological isoforms called prions. Of all human prion diseases, 10–15% of cases are genetic and the E200K mutation, which causes familial Creutz...

Cardiomyopathy is a co-morbidity of some prion diseases including genetic disease caused by mutations within the PrP gene (PRNP). Although the cellular prion protein (PrP) has been shown to protect against cardiotoxicity caused by oxidative stress, it is unclear if the cardiomyopathy is directly linked to PrP dysfunction. We differentiated cardiomy...

Human cerebral organoids are an exciting and novel model system emerging in the field of neurobiology. Cerebral organoids are spheres of self-organizing, neuronal lineage tissue that can be differentiated from human pluripotent stem cells and that present the possibility of on-demand human neuronal cultures that can be used for non-invasively inves...

Prion diseases are progressive, neurodegenerative diseases affecting humans and animals. Also known as the transmissible spongiform encephalopathies, for the hallmark spongiform change seen in the brain, these diseases manifest increased oxidative damage early in disease and changes in antioxidant enzymes in terminal brain tissue. Superoxide dismut...

La Crosse virus (LACV) is a mosquito-borne orthobunyavirus that causes approximately 60 to 80 hospitalized pediatric encephalitis cases in the United States yearly. The primary treatment for most viral encephalitis, including LACV, is palliative care, and specific antiviral therapeutics are needed. We screened the National Center for Advancing Tran...

The neuro-physiological properties of individuals with genetic pre-disposition to neurological disorders are largely unknown. Here we aimed to explore these properties using cerebral organoids (COs) derived from fibroblasts of individuals with confirmed genetic mutations including PRNPE200K, trisomy 21 (T21), and LRRK2G2019S, which are associated w...

Background Neurodegenerative diseases are highly complex making them challenging to model in cell culture. All cell types of the brain have been implicated as exerting an effect on pathogenesis, and disease progression is likely influenced by the cross-talk between the different cell types. Sophisticated investigation of the cellular level conseque...

Creutzfeldt–Jakob Disease (CJD) is a fatal, currently incurable, neurodegenerative disease. The search for candidate treatments would be greatly facilitated by the availability of human cell-based models of prion disease. Recently, an induced pluripotent stem cell derived human cerebral organoid model was shown to take up and propagate human CJD pr...

Cerebral organoids (COs) are a self-organizing three-dimensional brain tissue mimicking the human cerebral cortex. COs are a promising new system for modelling pathological features of neurological disorders, including prion diseases. COs expressing normal prion protein (PrPC) are susceptible to prion infection when exposed to the disease isoforms...

Microglia act as the protective immune cell of the brain. By surveying the tissue to identify and rectify problems, they function to maintain the health of brain cells. The prion protein N-terminal cleavage fragment, N1, has demonstrated neuroprotective activities in vitro and in vivo. This study aimed to elucidate whether N1 could modulate microgl...

α-synuclein strains may be responsible for the differing disease etiologies of the α-Synucleinopathies.

Background: La Crosse virus (LACV) is the leading cause of pediatric arboviral encephalitis in the USA. LACV encephalitis can result in learning and memory deficits, which may be due to infection and apoptosis of neurons in the brain. Despite neurons being the primary cell infected in the brain by LACV, little is known about neuronal responses to...

Prion diseases are transmissible and fatal neurological disorders associated with the misfolding of cellular prion protein (PrPC) into disease-causing isoforms (PrPD) in the central nervous system. The diseases have three etiologies; acquired through exposure to the infectious PrPD, sporadic, arising from no known cause, and hereditary due to famil...

Combination therapy may offer a way to slow the progression of prion diseases.

Neurons from the olfactory mucosa in combination with a highly sensitive amplification assay may offer a monitoring paradigm for α-synucleinopathies.

The prion protein functions as a carrier for delivering Aβ to exosomes during Alzheimer’s disease.

Abstract For the transmissible, neurogenerative family of prion diseases, few human models of infection exist and none represent structured neuronal tissue. Human cerebral organoids are self-organizing, three-dimensional brain tissues that can be grown from induced pluripotent stem cells. Organoids can model aspects of neurodegeneration in Alzheime...

α-synuclein oligomer uptake by neurons and oligodendrocytes exploits connexin-32.

The brain lymphatic system contributes to TAU clearance in mice.

Neurogenesis continues in the post-developmental brain throughout life. The ability to stimulate the production of new neurones requires both quiescent and actively proliferating pools of neural stem cells (NSCs). Actively proliferating NSCs ensure that neurogenic demand can be met, whilst the quiescent pool makes certain NSC reserves do not become...

α-Synuclein seeding activity can be measured quickly and might prove useful for the diagnosis of α-synucleinopathies.

In vivo near-infrared (NIR) imaging of molecular processes at the preclinical stage promises to provide more valuable mechanistic information about pathological pathways involved in neurodegeneration. NIR imaging has the potential to improve in vivo therapeutic screening protocols by enabling noninvasive monitoring of presymptomatic responses to tr...

Traditional primary and secondary cell cultures have been used for the investigation of prion biology and disease for many years. While both types of cultures produce highly valid and immensely valuable results, they also have their limitations; traditional cell lines are often derived from cancers, therefore subject to numerous DNA changes, and pr...

The ability to model brain tissue in three-dimensions offers new potential for elucidating functional cellular interactions and corruption of such functions during pathogenesis. Many protocols now exist for growing neurones in three-dimensions and these vary in complexity and cost. Herein, we describe a straight-forward method for generating three-...

Eight-hydroxyquinolines (8HQs) are a class of compounds that have been identified as potential therapeutics for a number of neurodegenerative diseases. Understanding the influence of structural modifications to the 8HQ scaffold on cellular behaviour will aid the identification of compounds that might be effective in treating dementias. In this stud...

The prion protein (PrP), through misfolding, is widely known for its causative role in prion diseases, which are transmissible neurodegenerative diseases of humans and animals. There is still no defined function assigned to PrP, especially in the central nervous system, despite many studies in this area. Proposed functions are protean and include s...

Accumulation of the β-amyloid (Aβ) peptide in extracellular senile plaques rich in copper and zinc is a defining pathological feature of Alzheimer's disease (AD). The Aβ1-x (x=16/28/40/42) peptides have been the primary focus of Cu(II) binding studies for more than 15 years; however, the N-truncated Aβ4-42 peptide is a major Aβ isoform detected in...

Accumulation of the β-amyloid (Aβ) peptide in extracellular senile plaques rich in copper and zinc is a defining pathological feature of Alzheimer′s disease (AD). The Aβ1–x (x=16/28/40/42) peptides have been the primary focus of CuII binding studies for more than 15 years; however, the N-truncated Aβ4–42 peptide is a major Aβ isoform detected in bo...

Internal cleavage of the cellular prion protein generates two well characterised N-terminal fragments, N1 and N2. These fragments have been shown to bind to anionic phospholipids at low pH. We sought to investigate binding with other lipid moieties and queried how such interactions could be relevant to the cellular functions of these fragments. Bot...

The protein misfolding cyclic amplification (PMCA) technique has become a widely-adopted method for amplifying minute amounts of the infectious conformer of the prion protein (PrP). PMCA involves repeated cycles of 20 kHz sonication and incubation, during which the infectious conformer seeds the conversion of normally folded protein by a templating...

The beta-amyloid (Aβ) peptide and the Aβ-oligomer receptor, prion protein (PrP), both influence neurogenesis. Using in vitro murine neural stem cells (NSCs), we investigated whether Aβ and PrP interact to modify neurogenesis. Aβ imparted PrP-dependent changes on NSC self-renewal, with PrP-ablated and wild-type NSCs displaying increased and decrease...

The prion protein (PrP), widely recognized to misfold into the causative agent of the transmissible spongiform encephalopathies, has previously been shown to bind to lipid membranes with binding influenced by both membrane composition and pH. Aside from the misfolding events associated with prion pathogenesis, PrP can undergo various posttranslatio...

The prion protein (PrP(C)) when mis-folded is causally linked with a group of fatal neurodegenerative diseases called transmissible spongiform encephalopathies or prion diseases. PrP(C) normal function is still incompletely defined with such investigations complicated by PrP(C) post-translational modifications, such as internal cleavage, which feas...

The compound 4-{[(4-hydroxyphenyl)amino]methyl}-2,6-di-tert-butylphenol (TRS1) was developed using a “mix-and-match” drug design approach for antioxidant-based neuroprotection and therapy. The commonly-used antipyretic drug, paracetamol, which is also found to act as a mild anti-inflammatory, was further modified by attaching a radical scavenger mo...

Oxidative stress as a contributor to neuronal death during prion infection is supported by various oxidative damage markers accumulating in the brain during the course of disease. The cellular substrate of the causative agent, the prion protein, is also linked with protective functions against oxidative stress. Our previous work has found that in c...

Prion disease transmission and pathogenesis are linked to misfolded, typically protease resistant (PrPres) conformers of the normal cellular prion protein (PrPC), with the former posited to be the principal constituent of the infectious 'prion'. Unexplained discrepancies observed between detectable PrPres and infectivity levels exemplify the comple...

Figure S2. Localisation of PrP, organelle and membrane markers in Nycodenz density gradient fractions of alternative cell lines. Representative immunoblots of subcellular fractions obtained from vector only transfected RK13 cells (vecRK13), GT1-7H and (M1000 infected) GT1-7H-inf, subject to SDS-PAGE and western blotting as described in the methods....

Figure S4. Illustration of the reduction of infectious titre in a 35-day extension of incubation period in Tga20 mice. Regression analysis (modelled on the results of an incubation time interval assay based on quantal end-point dose titration of M1000 brain homogenate in Tga20 mice) was used to plot the relationship between incubation period and ti...

Figure S7. Exogenous cellular co-factors from vecRK13 do not differentially increase the efficiency of M1000 infection in vitro. (A) Cell blot showing PrPres levels produced by recipient MoRK13 cells exposed to M1000 brain homogenate diluted to three different final concentrations (as indicated) with either 1:4 lysis buffer (LB):medium or 1:4 vecRK...

Figure S3. Localisation of PrP isoforms, organelle and membrane markers in Nycodenz density gradient fractions of M1000 brain. Representative immunoblots of subcellular fractions obtained from terminal M1000 brain. Brains were homogenised in a comparative way to the detergent-free cell lysis described in the methods, and then subject to Nycodenz gr...

Figure S6. PrPres profiles in Tga20 mice inoculated with 0.01% (w/v) M1000 brain homogenate, MoRK13-inf and selected MoRK13-inf subcellular fractions. Representative western blot (A) and quantification (B) of PrPres glycoform ratios in terminal mice brains after intracerebral inoculation as indicated. (A) 10 μl of PK digested (100 μg/ml final conce...

Figure S8. Lesion profiles from Tga20 mice intracerebrally inoculated with M1000 brain homogenate diluted in PBS and high density Nyocodenz with or without MoRK13 cell lysate content. Lesion profiles generated by quantification of the degree of vacuolation (H&E), reactive astrocytosis (GFAP) and PrP plaque deposition in each brain region (HI - hipp...

Figure S1. Disparity in PrPres levels and relative infectivity in the MoRK13-inf cell prion infection model. Representative western blot (A) and quantification (B) of PrPres levels in MoRK13-inf whole cell lysates relative to M1000 brain homogenate (n = 3). Samples were balanced for total protein; 5 μg total protein was resolved in untreated (-) la...

Figure S5. Assessment of neuropathology in the brains of Tga20 mice inoculated with 0.01% (w/v) M1000 brain homogenate, MoRK13 and MoRK13-inf whole cell lysates and selected MoRK13-inf fractions. Representative photomicrographs demonstrating the degree of (A) vacuolation in hematoxylin and eosin-stained (H & E) sections, (B) astrocytic gliosis in g...

Neuronal loss is a pathological feature of prion diseases for which increased reactive oxygen species (ROS) and consequent oxidative stress is one proposed mechanism. The processes underlying ROS production in prion disease and the precise relationship to misfolding of the prion protein remain obscure. Using cell culture models of prion infection w...