摘要:
The Applicants have established a simple, rapid assay of measuring the dynamics of self-assembling systems of biological molecules, based on stable isotope labeling technology that can be used in intact animals including humans. Examples of self-assembling systems of biological molecules include microtubule polymers, actin filaments, amyloid-beta plaques or fibrils, prion plaques or fibrils, fibrin aggregates, tau filaments (e.g., neurofibrillary tangles), α-synuclein filaments, and mutant hemoglobin aggregates. The method reveals constitutive differences in the dynamics of assembly and disassembly between tissues and is sensitive to the action of compounds that stabilize these dynamics.
摘要:
The invention disclosed herein describes a novel therapeutic target for motoneuron diseases (altered dynamics of microtubules in neurons); a method for measuring the state of activity of this therapeutic target in subjects with established, incipient, or potential motoneuron disease; the discovery of drug agents that modulate neuronal microtubule dynamics in living subjects with motoneuron diseases; the discovery that administration of such agents, alone or in combinations, can provide marked neuroprotective therapy for living subjects with motoneuron diseases including delay in symptoms and prolongation of survival; and the discovery that monitoring of neuronal microtubule dynamics in subjects with motoneuron diseases, in response to therapeutic interventions, allows diagnostic monitoring for optimization of therapeutic regimen and strategy for individual subjects or for drug trials.
摘要:
Provided herein are methods for measuring a molecular flux rate based on analysis of isotopologue abundance within a mass isotopomer, e.g., using a high resolution mass spectrometric measurement. Such methods may be used, inter alia, to calculate a fraction of newly synthesized target molecules of interest, a replacement rate of target molecules of interest, and/or a rate of breakdown or degradation of target molecules of interest, e.g., based on isotopologue relative abundance.
摘要:
The invention disclosed herein describes a novel therapeutic target for motoneuron diseases (altered dynamics of microtubules and microtubule-mediated axonal transport of cargo molecules in neurons), with or without dementia, and in dementia; methods for measuring the state of activity of this therapeutic target in subjects with established, incipient, or potential motoneuron disease, with or without dementia, and in dementia; the discovery of drug agents that modulate neuronal microtubule dynamics in living subjects with motoneuron diseases; the discovery that administration of such agents, alone or in combinations, can improve MT-mediated transport of cargo molecules along and through axons; the discovery that such modulation of altered microtubule dynamics and improvement in MT -transport of molecules along axons can provide marked neuroprotective therapy for living subjects with motoneuron diseases, including delay in symptoms and prolongation of survival; and the discovery that monitoring of microtubule-mediated axonal transport of cargo molecules in response to therapeutic interventions in subjects with motoneuron diseases, with or without dementia,and in dementia allows diagnostic monitoring, to optimize therapeutic regimens and treatment strategies in individual subjects or in drug trials.
摘要:
Provided herein are methods for measuring a molecular flux rate proteins and polypeptides based on analysis of isotopologue abundances of immonium ion fragments within a mass isotopomer using a high resolution mass spectrometric measurement. Such methods may be used, inter alia, to calculate a fraction of newly synthesized protein or polypeptide molecules of interest, a replacement rate of target molecules of interest, and/or a rate of breakdown or degradation of target molecules of interest, e.g., based on isotopologue relative abundances.
摘要:
Provided herein are method for measuring the rate of synthesis, breakdown, transport, or other kinetic parameters of a protein in a tissue of medical interest, without requiring physical sampling of the tissue, by a measurement of the protein in a body fluid. Methods may include selecting one or more target proteins in a tissue; administering an isotope-labeled molecule to a subject for a period of time sufficient for said isotope-labeled molecule to enter into and label the one or more target proteins to produce one or more isotope-labeled target proteins; collecting a volume of a body fluid, wherein the volume comprises one or more isotope-labeled target proteins that escaped or were released from the tissue; enriching or isolating the one or more isotope-labeled target proteins from the volume; performing a mass spectrometric measurement of the isotopic content, rate of incorporation, and/or pattern or rate of change in isotopic content and/or pattern of isotope labeling of the one or more enriched or isolated isotope-labeled target proteins; and calculating at least one kinetic parameter of the one or more enriched or isolated isotope-labeled target proteins, where the kinetic parameter of the one or more isotope-labeled target proteins from the volume of a body fluid reflects the corresponding kinetic parameter of the one or more target proteins in the tissue; and inferring the at least one kinetic parameter of the one or more target proteins in the tissue based on the corresponding at least one kinetic parameter of the one or more target proteins in the body fluid.
摘要:
The methods of the present invention allow for the measurement of proliferation, clonal expansion trafficking and/or recruitment of lymphocytes into lymphoid tissue in an in vivo setting. Proliferation, clonal expansion, recruitment and/or trafficking of lymphocytes are measured by using stable isotopes to label newly synthesized DNA, isolating the newly-labeled DNA, and quantifying enrichment of the isolated DNA with mass spectrometry or other appropriate techniques. Such methods are useful in screening candidate drugs for stimulatory or inhibitory effects on proliferation, clonal expansion, recruitment and/or trafficking of lymphocytes. The methods also allow for the discovery of therapeutic agents in disorders of immune regulation such as HIV infection and for optimizing vaccine efficacy.
摘要:
The present invention is based on the finding that enrichment of D3-creatinine in a urine sample following oral administration of a single defined dose of D3-creatine can be used to calculate total-body creatine pool size and total body skeletal muscle mass in a subject. The invention further encompasses methods for detecting creatinine and D3-creatinine in a single sample. The methods of the invention find use, inter alia, in diagnosing disorders related to skeletal muscle mass, and in screening potential therapeutic agents to determine their effects on muscle mass.