Project

Cryo-EM Structural Analysis of Cilia from PCD (Primary Ciliary Dyskinesia) Patients

Short Summary

We will analyze the 3D structure of motile cilia from human patients with primary ciliary dyskinesia (PCD) using cryo-electron tomography. With this approach, we will see how genetic defects of various ciliary proteins result in various symptoms of PCD. Specimen obtained from biopsies are cultured, embedded in amorphous ice, and measured by electron microscopy. We reconstruct the 3D structure of cilia from electron micrographs, clarify the defect of protein network and reveal which molecules are the causes of this disease.

Goals

The aim of this project is to establish a way to analyze 3D structure of motile cilia from patients by cryo-electron tomography and establish structure-based diagnosis of PCD.

Significance

Cryo-electron microscopy provides a wide view of 3D information of motile cilia at near atomic resolution. We expect that cryo-EM will enable diagnosis of cases of PCD, which were difficult to diagnose in the past with traditional methods, such as EM of thin sections.

Background

Motile cilia are cellular organelles, which make a beating motion to enable cells to swim or to generate extracellular fluid flow. In humans, motile cilia function in the respiratory system, the oviduct, the brain and in embryos. Defects in motile cilia cause various diseases (Primary Ciliary Dyskinesia, PCD). Since symptoms vary and there are a number of genes causing PCD, diagnosis of PCD is not straightforward and takes long time.