Stem Cell Sinoatrial Node Production via TBX and Myh6 Selection

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Solution Overview

Problem

Current methods for obtaining cardiac pacemaker cells, such as direct reprogramming and stem cell differentiation, face challenges in efficiency and purity, with existing approaches yielding low yields of functional sinoatrial node cells and lacking sensitivity to hormonal stimulation, leading to complications in treating sick sinus syndrome.

Innovation Solution

A method involving the introduction of a TBX transcription factor into stem cells, combined with an antibiotic selection using the Myh6 promoter, to differentiate stem cells into sinoatrial node cells with enhanced pacemaker properties, achieving high yields and functional maturity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If direct reprogramming of heart muscle cells is performed using viral vectors, then pacemaker cell transformation is achieved, but spatial and time-regulatable expression cannot be controlled and efficiency is very low

Engineering Contradiction:

Improvepacemaker cell transformationVSAvoidreprogramming efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The reprogramming process is divided into distinct stages: first introducing TBX transcription factor to initiate pacemaker gene expression, then applying antibiotic selection to purify the transformed cells. This segmentation allows control over both transformation reliability and selection efficiency.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent introduces the TBX transcription factor before applying antibiotic selection, preparing the cells in advance for the selection process. This preliminary action ensures that only cells that have successfully undergone reprogramming survive the antibiotic treatment, thereby increasing overall efficiency.

Inventive Principle:

Principle #10Preliminary action

2Adaptability or versatility

If stem cells are differentiated into pacemaker cells without selection, then cell diversity is maintained, but purity of functional sinoatrial node cells is low

Engineering Contradiction:

Improvecell differentiation capacityVSAvoidpacemaker cell purity

Core Design Contradiction:

Adaptability or versatilityVSManufacturing precision

Solution Approach 1:

The patent uses an antibiotic resistance gene as an intermediary marker linked to the Myh6 promoter. This intermediary allows indirect selection of pacemaker cells through antibiotic resistance, achieving high purity without directly selecting for pacemaker-specific markers during differentiation.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent replaces manual or physical selection methods with biochemical selection using antibiotic resistance. This substitution enables automated, high-throughput purification of pacemaker cells based on genetic markers rather than phenotypic characteristics.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Productivity

If existing stem cell differentiation methods are used, then cell production is achieved, but beat frequencies are too low and functional maturity is insufficient

Engineering Contradiction:

Improvecell production yieldVSAvoidfunctional maturity

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent changes the genetic parameters of the differentiated cells by introducing TBX transcription factors that specifically upregulate pacemaker gene programs. This parameter change transforms the functional characteristics of the cells, increasing beat frequencies to physiological ranges and achieving mature pacemaker functionality.

Inventive Principle:

Principle #35Parameter changes

4Reliability

If artificial pacemakers are implanted, then cardiac rhythm control is achieved, but patients face high risks of infection and battery discharge complications

Engineering Contradiction:

Improvecardiac rhythm controlVSAvoidinfection and battery discharge risks

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent creates biological copies of pacemaker cells from stem cells that can be transplanted into the patient's heart. These living cell copies replicate the pacemaker function naturally, eliminating the need for mechanical devices with batteries and reducing infection risks associated with implanted electronics.

Inventive Principle:

Principle #26Copying

Data Source

PatentUS11661582B2Method for producing sinoatrial node cells (pacemaker cells) from stem cells, and use of the produced sinoatrial node cells

Publication Date: 2023.05.30 UNIVERSITY OF ROSTOCK

AI summary

The electrical pacemakers currently being used for the therapeutic approaches for treatment of “sick sinus syndrome” are not hormonally regulatable and entail risks through infections or premature battery discharge. These problems could be overcome by means of “biological cardiac pacemakers” obtained from pluripotent stem cells (PSCs). It has been shown that the controlled differentiation of stem cells with TBX, inductors of sinoatrial node cells, and an additional Myh6 promoter-specific antibiotic selection can give cardiomyocyte aggregates consisting to an extent of more than 80% of physiologically functional pacemaker cells. These induced sinoatrial bodies (“iSABs”) for the first time exhibited very high beat frequencies (300-400 bpm), similar to those in a murine heart, and were able to stably rhythmically stimulate heart muscle cells ex vivo. In the iSAB transcriptome decoded by means of RNA-seq, it was possible to assign almost all the genes to the ontologies of heart function/heart development and the structures of contractile cells. Overall, this is the first example of a high-purity functional sinoatrial tissue derived from stem cells, which means that a crucial step for future cell therapy and the testing of medicaments in vitro is being implemented.