Partial Restoration of Mutant Enzyme Homeostasis in Three Distinct Lysosomal Storage Disease Cell Lines by Altering Calcium Homeostasis

A lysosomal storage disease (LSD) results from deficient lysosomal enzyme activity, thus the substrate of the mutant enzyme accumulates in the lysosome, leading to pathology. In many but not all LSDs, the clinically most important mutations compromise the cellular folding of the enzyme, subjecting it to endoplasmic reticulum–associated degradation instead of proper folding and lysosomal trafficking. A small molecule that restores partial mutant enzyme folding, trafficking, and activity would be highly desirable, particularly if one molecule could ameliorate multiple distinct LSDs by virtue of its mechanism of action. Inhibition of L-type Ca2+ channels, using either diltiazem or verapamil—both US Food and Drug Administration–approved hypertension drugs—partially restores N370S and L444P glucocerebrosidase homeostasis in Gaucher patient–derived fibroblasts; the latter mutation is associated with refractory neuropathic disease. Diltiazem structure-activity studies suggest that it is its Ca2+ channel blocker activity that enhances the capacity of the endoplasmic reticulum to fold misfolding-prone proteins, likely by modest up-regulation of a subset of molecular chaperones, including BiP and Hsp40. Importantly, diltiazem and verapamil also partially restore mutant enzyme homeostasis in two other distinct LSDs involving enzymes essential for glycoprotein and heparan sulfate degradation, namely α-mannosidosis and type IIIA mucopolysaccharidosis, respectively. Manipulation of calcium homeostasis may represent a general strategy to restore protein homeostasis in multiple LSDs. However, further efforts are required to demonstrate clinical utility and safety

The New NASA Orbital Debris Engineering Model ORDEM2000

The NASA Orbital Debris Program Office at Johnson Space Center has developed a new computer-based orbital debris engineering model, ORDEM2000, which describes the orbital debris environment in the low Earth orbit region between 200 and 2000 km altitude. The model is appropriate for those engineering solutions requiring knowledge and estimates of the orbital debris environment (debris spatial density, flux, etc.). ORDEM2000 can also be used as a benchmark for ground-based debris measurements and observations. We incorporated a large set of observational data, covering the object size range from 10 mm to 10 m, into the ORDEM2000 debris database, utilizing a maximum likelihood estimator to convert observations into debris population probability distribution functions. These functions then form the basis of debris populations. We developed a finite element model to process the debris populations to form the debris environment. A more capable input and output structure and a user-friendly graphical user interface are also implemented in the model. ORDEM2000 has been subjected to a significant verification and validation effort. This document describes ORDEM2000, which supersedes the previous model, ORDEM96. The availability of new sensor and in situ data, as well as new analytical techniques, has enabled the construction of this new model. Section 1 describes the general requirements and scope of an engineering model. Data analyses and the theoretical formulation of the model are described in Sections 2 and 3. Section 4 describes the verification and validation effort and the sensitivity and uncertainty analyses. Finally, Section 5 describes the graphical user interface, software installation, and test cases for the user