Yeast Cells Expressing ApoE Proteins for Toxicity Screening
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Solution Overview
Problem
Current methods lack effective ways to identify and address the toxicity induced by human Apolipoprotein E (ApoE) proteins, particularly ApoE4, which is linked to neurodegenerative diseases like Alzheimer's, as these proteins cause cellular dysfunction and are difficult to study directly in human cells.
Innovation Solution
Engineering yeast cells to express ApoE proteins, allowing for the screening of compounds and genetic factors that modulate ApoE-induced toxicity, using constructs with promoters linked to nucleic acids encoding ApoE proteins and signal sequences, enabling the identification of genetic suppressors or enhancers of toxicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If human ApoE proteins are expressed in yeast cells, then the ability to screen for compounds that modulate ApoE-induced toxicity is improved, but the growth and viability of the yeast cells deteriorate
Solution Approach 1:
The patent uses yeast cells as an intermediary model system to study human ApoE toxicity. The yeast cells express human ApoE proteins through genetically engineered expression constructs, allowing researchers to screen for compounds that modulate toxicity without directly working with human cells. This intermediary approach enables reliable screening while managing the toxicity challenge through controlled expression systems and multiple yeast strain options.
2Measurement precision
If ApoE proteins are expressed in yeast cells, then compounds that modulate ApoE-induced toxicity can be identified, but the yeast cells experience decreased growth or viability
Solution Approach 1:
The patent employs dynamic control of ApoE expression through inducible promoters in the yeast expression system. This allows researchers to control when and how much ApoE is expressed, enabling toxicity detection at specific time points while maintaining cell growth during other periods. The dynamic expression control optimizes both measurement precision for toxicity detection and productivity for maintaining sufficient cell populations.
3Adaptability or versatility
If yeast cells are engineered to express ApoE proteins, then screening assays can be performed to identify therapeutic agents, but the cellular dysfunction caused by ApoE complicates the system
Solution Approach 1:
The patent segments the screening system by using yeast cells that are genetically engineered to express only the specific human ApoE proteins of interest, rather than relying on yeast's native protein repertoire. This segmentation allows the system to be highly adaptable for screening different ApoE isoforms and compounds while maintaining reliability by using a simplified, controlled cellular background that reduces confounding cellular dysfunction from other sources.
Data Source
AI summary
Disclosed are yeast cells expressing a polypeptide comprising a signal sequence and a human ApoE protein. In some embodiments the polypeptide comprises ApoE2. In some embodiments the polypeptide comprises ApoE3. In some embodiments the polypeptide comprises ApoE4. Also disclosed are methods of screening yeast cells to identify compounds that prevent or suppress Apo-induced toxicity. Compounds identified by such screens can be used to treat or prevent neurodegenerative disorders such as Alzheimer's disease. Also disclosed are methods of screening yeast cells to identify genetic suppressors or enhancers of ApoE-induced toxicity. Also disclosed are genetic suppressors or enhancers of ApoE-induced toxicity identified using the methods, and human homologs thereof. Also disclosed are methods of identifying compounds that modulate expression or activity of genetic modifiers of ApoE-induced toxicity.