Extracellular Peptide-Ligand Dimerization Actuator Receptor Design for Reversible and Spatially Dosed 3D Cell-Material Communication DOI Creative Commons
Matthias Recktenwald,

Ritankar Bhattacharya,

Mohammed Mehdi Benmassaoud

et al.

ACS Synthetic Biology, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 20, 2024

Transmembrane receptors that endow mammalian cells with the ability to sense and respond biomaterial-bound ligands will prove instrumental in bridging fields of synthetic biology biomaterials. Materials formed thiol-norbornene chemistry are amenable thiol-peptide patterning, this study reports rational design reversibly activate cellular responses based on peptide-ligand recognition. This transmembrane receptor platform, termed Extracellular Peptide-ligand Dimerization Actuator (EPDA), consists stimulatory or inhibitory pairs come together upon extracellular peptide dimer binding corresponding monobody receptors. Intracellularly, Stimulatory EPDAs phosphorylate a substrate merges two protein halves, whereas Inhibitory revert split proteins back their unmerged, inactive state via dephosphorylation. To identify ligand-receptor pairs, over 2000 candidate monobodies were built silico using PETEI, novel computational algorithm we developed. The top 30 predicted affinity tested experimentally, induced highest change merging (green fluorescent protein, GFP) incorporated final EPDA design. In soluble form, peptides induce intracellular GFP time- concentration-dependent manner, varying levels green fluorescence observed dosing. EPDA-programmed encapsulated hydrogels patterned domains exhibited 3D activation deactivation location within peptide-patterned hydrogels. can recognize myriad peptide-ligands bound materials, beyond fluorescence, widely applicable biological research regenerative medicine.

Language: Английский

Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues DOI Creative Commons
Jeremy C. Tran, Christopher J. Kuffner, Alexander M. Marzilli

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 20, 2025

Synthetic Notch (SynNotch) receptors function like natural proteins and can be used to install customized sense-and-respond capabilities into mammalian cells. Here, we introduce an adaptor-based strategy for regulating SynNotch activity via fluorescein isomers analogs. Using optimized fluorescein-binding receptor, describe ways chemically control signaling, including approach based on a bio-orthogonal chemical ligation spatially controllable the photo-patterned uncaging of o-nitrobenzyl-caged conjugate. We further show that fluorescein-conjugated extracellular matrix (ECM)-binding peptides regulate depending folding state collagen-based ECM networks. To demonstrate utility these tools, apply them activate dose-dependent gene expression responses induce myogenic-like phenotypes in multipotent fibroblasts with spatiotemporal microenvironmental control. Overall, as versatile tool transcriptional ligands clinically-approved dye. endow cells custom capabilities. authors controlling diverse stimuli fluorescein-conjugates.

Language: Английский

Citations

0
Designer mammalian living materials through genetic engineering DOI
Mariana Gameiro, J. Almeida-Pinto, Beatriz S. Moura

et al.

Bioactive Materials, Journal Year: 2025, Volume and Issue: 48, P. 135 - 148

Published: Feb. 15, 2025

Language: Английский

Citations

0
Responsive DNA artificial cells for contact and behavior regulation of mammalian cells DOI Creative Commons
Miao Wang,

Hexin Nan,

Meixia Wang

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: March 11, 2025

Artificial cells have emerged as synthetic entities designed to mimic the functionalities of natural cells, but their interactive ability with mammalian remains challenging. Herein, we develop a generalizable and modular strategy engineer DNA-empowered stimulable artificial designated regulate (STARM) via contact-dependent communication. Constructed through temperature-controlled DNA self-assembly involving liquid-liquid phase separation (LLPS), STARMs feature organized all-DNA cytoplasm-mimic membrane-mimic compartments. These compartments can integrate functional nucleic acid (FNA) modules light-responsive gold nanorods (AuNRs) establish programmable sense-and-respond mechanism specific stimuli, such light or ions, orchestrating diverse biological functions, including tissue formation cellular signaling. By combining two designer into dual-channel system, achieve orthogonally regulated signaling in multicellular communities. Ultimately, vivo therapeutic efficacy STARM light-guided muscle regeneration living animals demonstrates promising potential smart regenerative medicine. Interaction between is major goal huge potential. Here, authors developed DNA-assembled, stimuli-responsive capable communication precisely modulate cell behaviors.

Language: Английский

Citations

0
Post-transcriptional modular synthetic receptors DOI
Xiaowei Zhang, Luis Santiago Mille, Kristjan Eerik Kaseniit

et al.

Nature Chemical Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 28, 2025

Language: Английский

Citations

0
Engineering of CD8 + T cells with an HIV-specific synthetic notch receptor to secrete broadly therapeutic antibodies for combining antiviral humoral and cellular immune responses DOI Creative Commons

Lina Meng,

Hongwei Zhao, Shuang Chang

et al.

mBio, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

ABSTRACT The application of immunotherapeutic strategies, such as chimeric antigen receptor-T cells and broadly neutralizing antibodies (bNAbs), for the treatment human immunodeficiency virus (HIV) infection is hindered by latent reservoirs viral escape. Achieving long-term control load in absence antiretroviral therapy requires a combination approach utilizing these strategies. For this purpose, we developed novel anti-HIV-1 synthetic Notch (synNotch) cells, termed CD4-17b-VN, which express both bNAb (VRC01) bispecific T cell-engaging protein (N6-αCD3) with antigenic control. synNotch receptor-expressing can sense presented on HIV-1 particles surface target cells. A cell line equipped CD4-17b-VN circuit could effectively VRC01 N6-αCD3 secretion upon sensitization, suppress diverse subtypes strains, mediate specific bypass cytotoxic activity against infected latency-reactivated Additionally, CD8 + exhibited long-lasting suppression stronger killing effect vitro . Importantly, demonstrated that receptor did not increase susceptibility to engineered Our study validates concept platform-based therapeutic deliver an antigen-controlled manner, may have important implications functional cure AIDS. IMPORTANCE Adoptive transfer effector modified has been proposed applicable treat infection. (SNR) system serves versatile tool, enabling customized programming input output functions mammalian Herein, report engineering targeting cell-free coupling antibody neutralization cytotoxicity. findings demonstrate CD4-17b SNR enables controllable production recognition particle antigens bifunctional synNotch-T Human primary replication reduce , without undesired risk being virus, suggesting their potential candidacy AIDS prospects future clinical applications.

Language: Английский

Citations

0
Induced Neural Progenitor Specification from Human Pluripotent Stem Cells by a Refined Synthetic Notch Platform DOI Creative Commons

Catherine A. Hamann,

Andrew Kjar, Hyosung Kim

et al.

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: May 6, 2025

Historically, studying the development of brain and central nervous system (CNS) tissues has been challenging. Human pluripotent stem cell (hPSC) technology allowed for in vitro reconstitution relevant, early trajectories by using small molecules recombinant proteins to guide differentiation cells toward relevant CNS phenotypes. However, many these protocols fail recapitulate cell-guided programs intrinsic embryonic development, particularly signaling centers that emerge within neural tube during formation. Located on ventral end tube, floor plate acts as one such center pattern dorsal/ventral axis secreting morphogen Sonic Hedgehog (SHH). Here, we present a method synthetic Notch (synNotch) receptor platform regulate SHH production subsequent fate specification. We show widely used configuration orthogonal synNotch ligand green fluorescent protein (GFP) mounted platelet-derived growth factor receptor-β transmembrane chassis does not allow robust artificial synNotch-hPSCs ("receivers") cocultured with ligand-presenting hPSCs ("senders"). discovered refined designs membrane-bound GFP-ligand efficient activation hPSC receivers. A variant this enhanced drives sender:hPSC receiver cocultures gives rise plate-like types seen development. This revised potential morphogenesis studies designed uncover key paradigms human

Language: Английский

Citations

0
Templated Pluripotent Stem Cell Differentiation via Substratum-Guided Artificial Signaling DOI Creative Commons

Hannah J. Brien,

Joanne C. Lee, Jhanvi Sharma

et al.

ACS Biomaterials Science & Engineering, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 1, 2024

The emerging field of synthetic morphogenesis implements biology tools to investigate the minimal cellular processes sufficient for orchestrating key developmental events. As continues grow, there is a need new that enable scientists uncover nuances in molecular mechanisms driving cell fate patterning emerge during morphogenesis. Here, we present platform combines engineering with biomaterial design potentiate artificial signaling pluripotent stem cells (PSCs). This platform, referred as PSC-MATRIX, extends use programmable biomaterials PSCs competent activate morphogen production through orthogonal signaling, giving rise opportunity probe events by initiating morphogenetic programs spatially constrained manner non-native channels. We show PSC-MATRIX enables temporal and spatial control transgene expression response bulk, soluble inputs Notch (synNotch)-engineered human an extended culture up 11 days. Furthermore, used regulate multiple differentiation via material-mediated engineered using ligand green fluorescent protein, highlighting potential this probing guiding acquisition. Overall, offers approach interrogate PSC could be applied variety protocols.

Language: Английский

Citations

0
Extracellular Peptide-Ligand Dimerization Actuator Receptor Design for Reversible and Spatially Dosed 3D Cell-Material Communication DOI Creative Commons
Matthias Recktenwald,

Ritankar Bhattacharya,

Mohammed Mehdi Benmassaoud

et al.

ACS Synthetic Biology, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 20, 2024

Transmembrane receptors that endow mammalian cells with the ability to sense and respond biomaterial-bound ligands will prove instrumental in bridging fields of synthetic biology biomaterials. Materials formed thiol-norbornene chemistry are amenable thiol-peptide patterning, this study reports rational design reversibly activate cellular responses based on peptide-ligand recognition. This transmembrane receptor platform, termed Extracellular Peptide-ligand Dimerization Actuator (EPDA), consists stimulatory or inhibitory pairs come together upon extracellular peptide dimer binding corresponding monobody receptors. Intracellularly, Stimulatory EPDAs phosphorylate a substrate merges two protein halves, whereas Inhibitory revert split proteins back their unmerged, inactive state via dephosphorylation. To identify ligand-receptor pairs, over 2000 candidate monobodies were built silico using PETEI, novel computational algorithm we developed. The top 30 predicted affinity tested experimentally, induced highest change merging (green fluorescent protein, GFP) incorporated final EPDA design. In soluble form, peptides induce intracellular GFP time- concentration-dependent manner, varying levels green fluorescence observed dosing. EPDA-programmed encapsulated hydrogels patterned domains exhibited 3D activation deactivation location within peptide-patterned hydrogels. can recognize myriad peptide-ligands bound materials, beyond fluorescence, widely applicable biological research regenerative medicine.

Language: Английский

Citations

0