Chloride Ion Biosensors for Human iPSC Models
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Solution Overview
Problem
Current methods for studying intracellular chloride levels often rely on non-representative cell models, such as rodent cells or immortalized cell lines, which may not accurately reflect human cellular responses, limiting the effectiveness of bioactive agent screening and therapeutic evaluation.
Innovation Solution
Development of chloride ion biosensors that can be targeted to specific intracellular locations, allowing for 2D or 3D imaging of human iPSC-derived cell models, enabling precise detection of chloride ion concentrations and fluctuations in membrane potentials, and can be configured for various cell types, including dopaminergic neurons and cardiomyocytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If non-representative cell models (rodent cells or immortalized cell lines) are used for studying intracellular chloride levels, then research can be conducted with available cell lines, but the results do not accurately reflect human cellular responses
Solution Approach 1:
The patent uses human induced pluripotent stem cells (iPSCs) to create accurate copies of human cellular systems. By reprogramming human somatic cells into pluripotent stem cells and then differentiating them into target cell types, the invention creates in vitro models that faithfully replicate human cellular behavior, thereby resolving the contradiction between measurement accuracy and human cell applicability
Solution Approach 2:
The invention changes the fundamental parameter of cell origin and type by using human iPSCs instead of rodent or immortalized cell lines. This parameter change transforms the cellular model from non-representative to representative of human physiology, enabling accurate measurement of chloride levels and cellular responses in human-relevant systems
2Measurement precision
If chloride ion biosensors are developed for specific intracellular locations, then precise detection of chloride concentrations is achieved, but the device complexity increases
Solution Approach 1:
The chloride ion biosensor is segmented into distinct functional domains: a chloride-sensing domain (such as a chloride channel or transporter), a linker region, and a reporter domain (fluorescent protein). This segmentation allows each component to perform its specific function while maintaining overall sensor functionality, achieving precise chloride detection without excessive complexity
Solution Approach 2:
The patent uses a linker region as an intermediary between the chloride-sensing domain and the fluorescent reporter domain. This intermediary element transmits conformational changes from the sensing domain to the reporter domain, enabling signal transduction while maintaining structural flexibility and avoiding overly complex direct connections
3Reliability
If human iPSC-derived cell models are used for drug screening, then representative human cellular responses are obtained, but the screening process becomes more complex
Solution Approach 1:
The patent performs preliminary differentiation of human iPSCs into target cell types (such as neurons, cardiomyocytes, or hepatocytes) before drug screening. This preliminary action creates stable, differentiated cell models that maintain human-specific physiological characteristics, thereby improving reliability for predicting human responses while establishing a reproducible workflow that manages complexity
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
These biosensors facilitate accurate screening and evaluation of candidate compounds for chemical, biological, and toxicological effects in humans and other mammals by providing representative models of human cellular responses, enhancing the predictive power of drug candidate assessments.
Implementation Method 1
The reporter domain is preferably a fluorescent reporter domain. The fluorescent reporter may fluoresce at wavelengths in the range from 500 nm to 1400 nm, and preferably has a fluorescent emission range of 500 nm-700 nm and a fluorescent excitation range of 400 nm-700 nm.
Implementation Method 2
The transmembrane domain is preferably adapted to integrate into a membrane of a human cell.
Data Source
AI summary
This invention provides biosensors, cell models, and methods of their use for monitoring chloride ion, where the biosensors can include targeting domains, sensing domains and reporting domains. Biosensors can be introduced into cells reprogrammed to represent experimental or pathologic cells of interest, including as detectors of chloride ions, as TempoChloro™ accomplishes.