Nihat, Akin; (2025) Developing tools to investigate human prion diseases. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Human prion diseases are universally fatal neurodegenerative conditions united by the autocatalytic, templated misfolding of the constitutive prion protein (PrP). This thesis presents work aimed at developing key cell-based and clinical tools to support their investigation. The study of human prions has been considerably impeded by the absence of a robust cell-based assay capable of reproducibly propagating and quantifying bona fide human prion infectivity, which has been a goal of the field for several decades. To establish such a system, we pursued a strategy of silencing PrP expression in murine CAD5 cells that have wide non-human prion tropism, followed by reconstitution with human PrP. In the first portion of this thesis (Chapters 3 and 4), we describe dividing CAD5 cells expressing human PrP that reproducibly accumulate PrPSc after exposure to sporadic Creutzfeldt-Jakob Disease (sCJD) and variant CJD (vCJD)-infected frontal cortex. sCJD-susceptible cells were further developed to i) increase cell sensitivity to prion infection through iterative single cell cloning, ii) confirm cellular propagation of de novo human prion infectivity via transmission to mouse bioassay, iii) create a robust human prion assay (HPA) with comparable sensitivity to mouse endpoint titration iv) derive cells with persistent sCJD infection that can be cured and adapted for therapeutic screening. Chapter 5 addresses the need for improved tools to objectively assess disease progression in sCJD, to support patient stratification and determine treatment response in clinical trials, and in care planning. The existing MRC Prion Disease Rating Scale (MRC Scale) provides a validated outcome measure for overall functional progression – however, given the wide heterogeneity in clinical phenotype and disease course, additional tools assessing specific functions would be of use. We describe the creation of complementary clinimetric examination scales to assess progressive cognitive and motor dysfunction in sCJD, using Rasch modelling. Together, these novel cell-based and clinical tools provide means to investigate previously inaccessible aspects of human prion disease.

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