TropR

Stages

1. 1.
   TropR receptor engineering(library_design)

   This stage creates the sensing architecture needed to convert cTnI detection into intracellular signaling and gene-expression control.

   Selection: Design cTnI-sensing chimeric receptors using extracellular scFvs and alternative intracellular signaling domains.

2. 2.
   Functional confirmation of cTnI-dependent signaling(functional_characterization)

   This stage verifies that the receptor works in relevant mammalian cell contexts before building therapeutic output lines.

   Selection: Confirm cTnI-dependent TropR functionality and synthetic-promoter control in HEK-derived cell lines and iPSC-derived cardiomyocytes.

3. 3.
   Construction of therapeutic monoclonal cell lines(library_build)

   This stage converts the sensing module into a therapeutic closed-loop cell product.

   Selection: Build monoclonal cell lines for cTnI-induced tenecteplase secretion with a doxycycline-triggered off-switch.

4. 4.
   Lead clone selection(hit_picking)

   This stage narrows multiple monoclonal lines to a lead clone with sensitivity matched to human AMI-relevant biomarker levels.

   Selection: Select a clone optimized to detect human AMI-relevant cTnI levels.

5. 5.
   Ex vivo thrombolytic validation(confirmatory_validation)

   This stage confirms that the selected therapeutic clone performs the intended closed-loop function in a clot-lysis assay.

   Selection: Test whether alginate-microencapsulated CardioProtect cells lyse fibrin clots in an ex vivo blood culture system under cTnI induction and doxycycline repression.

Steps

1. 1.
   Design cTnI-sensing TropR variantsengineered receptor

   Create a chimeric receptor that senses cTnI and couples detection to intracellular signaling.

   A sensing module is required before downstream gene-expression control and therapeutic output can be engineered.

2. 2.
   Confirm cTnI-dependent TropR function in mammalian cellssensing construct under test

   Verify that TropR drives synthetic-signaling-specific promoter outputs in response to cTnI.

   Functional confirmation is needed before investing in therapeutic clone construction.

3. 3.
   Construct monoclonal cTnI-inducible TNK-secreting cell lines with doxycycline off-switchtherapeutic cell construct

   Build therapeutic cell lines that convert cTnI sensing into tenecteplase secretion while retaining external shutoff control.

   Therapeutic output is added after receptor function is established.

4. 4.
   Select CardioProtect as the lead cloneselected lead clone

   Choose the monoclonal line with sensitivity optimized for human AMI-relevant cTnI levels.

   A single optimized lead clone is needed before confirmatory thrombolysis validation.

5. 5.
   Validate alginate-microencapsulated CardioProtect in ex vivo clot lysis assayencapsulated therapeutic cell product

   Test whether the selected clone performs strict cTnI-inducible, doxycycline-repressible thrombolysis.

   Confirmatory efficacy testing follows lead-clone selection to show the full closed-loop therapeutic behavior.