A Light‐Controlled TLR4 Agonist and Selectable Activation of Cell Subpopulations

Stutts, Lalisa; Esser‐Kahn, Aaron P.

doi:10.1002/cbic.201500164

Cited by 12 publications

(14 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…13 In addition, the indole nitrogen has been functionalized with nitroveratryloxycarbonyl (NVOC) to afford a light-activated TLR4 agonist, showing that this site is important for ligand binding despite docking studies suggesting it may be solvent accessible. 108 More recently, a further optimized pyramido- [5,4-b]indole TLR4 agonist (Compound 26, EC 50 : 600 nM, Fig. 4B) was shown to improve and bias human TLR4 activation over mouse TLR4 activation.…”

Section: ø Tlr4mentioning

confidence: 99%

Toll-like Receptor Agonist Conjugation: A Chemical Perspective

et al. 2018

Self Cite

View full text Add to dashboard Cite

Toll-like receptors (TLRs) are vital elements of the mammalian immune system that function by recognizing pathogen-associated molecular patterns (PAMPs), bridging innate and adaptive immunity. They have become a prominent therapeutic target for the treatment of infectious diseases, cancer, and allergies, with many TLR agonists currently in clinical trials or approved as immunostimulants. Numerous studies have shown that conjugation of TLR agonists to other molecules can beneficially influence their potency, toxicity, pharmacokinetics, or function. The functional properties of TLR agonist conjugates, however, are highly dependent on the ligation strategy employed. Here, we review the chemical structural requirements for effective functional TLR agonist conjugation. In addition, we provide similar analysis for those that have yet to be conjugated. Moreover, we discuss applications of covalent TLR agonist conjugation and their implications for clinical use.

show abstract

Section: ø Tlr4mentioning

confidence: 99%

Toll-like Receptor Agonist Conjugation: A Chemical Perspective

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…The fast response of endocytosis after illumination indicated that the behavior of the cells could be temporally controlled using the photocaged TLR8 model. The spatial control of TLR can also be measured by detecting the behavior of downstream NF‐κB [5d] . To test spatial constraints, we measured nuclear translocation of the p65 subunit of NF‐κB by stimulating a portion of the cells in a 20‐mm diameter well by using a 5‐mm diameter circular photo‐mask.…”

Section: Figurementioning

confidence: 99%

Photoactivation of Innate Immunity Receptor TLR8 in Live Mammalian Cells by Genetic Encoding of Photocaged Tyrosine

et al. 2021

View full text Add to dashboard Cite

The effectiveness of innate immune responses relies on an intricate balance between activation and regulation. TLR8, a member of the Toll‐like receptor (TLR) family, plays a fundamental role in host defense by sensing viral single‐stranded RNAs (ssRNAs). However, the molecular recognition and regulatory mechanism of TLR8 is not fully understood, especially in a whole‐cell environment. Here, we engineer the first light‐controllable TLR8 model by genetically encoding a photocaged tyrosine, NBY, into specific sites of TLR8. In the caged forms, the activity of TLR8 is masked but can be restored upon decaging by exposure to UV light. To explain the mechanism clearly, we divide the sites with light responsiveness into three groups. They can separately block the ligands that bind to the pockets of TLR8, change the interaction modes between two TLR8 protomers, and interfere with the interactions between TLR8 cytosolic domains with its downstream adaptor. Specifically, we use this chemical caging strategy to probe and evaluate the function of several tyrosine sites located at the interface of TLR8 homodimers with a previously unknown regulatory mode, which may provide a new strategy for TLR8 modulator development. Effects on downstream signaling pathways are monitored at the transcriptional and translational levels in various cell lines. By photoactivating specific cells within a larger population, this powerful tool can provide novel mechanistic insights, with potential in biotechnological and pharmaceutical applications.

show abstract

“…Microfluidic delivery is complementary to existing methods for local stimulation of intact lymphoid tissue. For instance, light-induced stimulation of photo-activated receptors 11,12,52 and release of caged molecules 53,54 offer potentially higher spatial and temporal resolution than fluidic delivery, but require genetically modified animals 55 or specifically designed cage systems for each molecule of interest. Microinjection is another well-established approach for local stimulation in brain 13,56 , although it may increase the risk of local tissue damage around the insertion site.…”

Section: Local Activation Of Immune Cells On-chipmentioning

confidence: 99%

Spatially resolved microfluidic stimulation of lymphoid tissue ex vivo

Ross

Belanger

Woodroof

et al. 2017

Analyst

View full text Add to dashboard Cite

The lymph node is a structurally complex organ of the immune system, whose dynamic cellular arrangements are thought to control much of human health. Currently, no methods exist to precisely stimulate substructures within the lymph node or analyze local stimulus-response behaviors, making it difficult to rationally design therapies for inflammatory disease. Here we describe a novel integration of live lymph node slices with a microfluidic system for local stimulation. Slices maintained the cellular organization of the lymph node while making its core experimentally accessible. The 3-layer polydimethylsiloxane device consisted of a perfusion chamber stacked atop stimulation ports fed by underlying microfluidic channels. Fluorescent dextrans similar in size to common proteins, 40 and 70 kDa, were delivered to live lymph node slices with 284 ± 9 μm and 202 ± 15 μm spatial resolution, respectively, after 5 s, which is sufficient to target functional zones of the lymph node. The spread and quantity of stimulation were controlled by varying the flow rates of delivery; these were predictable using a computational model of isotropic diffusion and convection through the tissue. Delivery to two separate regions simultaneously was demonstrated, to mimic complex intercellular signaling. Delivery of a model therapeutic, glucose-conjugated albumin, to specific regions of the lymph node indicated that retention of the drug was greater in the B-cell zone than in the T-cell zone. Together, this work provides a novel platform, the lymph node slice-on-a-chip, to target and study local events in the lymph node and to inform the development of new immunotherapeutics.

show abstract

A Light‐Controlled TLR4 Agonist and Selectable Activation of Cell Subpopulations

Cited by 12 publications

References 34 publications

Toll-like Receptor Agonist Conjugation: A Chemical Perspective

Toll-like Receptor Agonist Conjugation: A Chemical Perspective

Photoactivation of Innate Immunity Receptor TLR8 in Live Mammalian Cells by Genetic Encoding of Photocaged Tyrosine

Spatially resolved microfluidic stimulation of lymphoid tissue ex vivo

Contact Info

Product

Resources

About