Multifunctional Cell Research Articles

Structural basis for the molecular recognition of various RGD-containing ligands by integrin αVβ3: The role of the specificity-determining loop (SDL).

Integrins are multifunctional cell surface receptors that play critical roles in normal physiological processes such as proliferation, survival, migration, and cell fate transitions. Integrins are implicated in several pathological conditions, including inflammation, tumor metastasis, lung fibrosis, and autoimmune disorders. Recent studies show that apart from their endogenous ligands, αVβ3 and α5β1 integrins bind directly to the hydroxylated form of cholesterol (25HC and 24HC) to upregulate pro-inflammatory response. Among the integrins, αVβ3 integrin has been considered a viable target for treating many diseases. Despite enormous efforts, currently available RGD-based anti-integrin drugs lack specificity, exhibit partial agonism, and cause serious side effects, including tumor growth and angiogenesis. The development of safe and efficient inhibitors requires a thorough understanding of the structural basis for outside-in activation and inhibition mechanism of integrin αvβ3. In this study, we performed a comprehensive residue-residue contact and network analysis of the αvβ3 integrin headpiece using all available experimental structures. Surprisingly, our analyses show that specific orientation and interactions of residues from the specificity-determining loop (SDL) are critical in the molecular recognition and regulation of integrin αvβ3. In addition, network analysis reveals that residues from the allosteric site (site II) connect to the primary RGD-binding site via SDL, which acts as an interface for the allosteric modulation. The obtained results provide a comprehensive structural perspective, useful insights into the distinct features of active and inactive conformations, and the role of SDL in the ligand recognition process and allostery.

Pericytes control vascular stability and auditory spiral ganglion neuron survival.

The inner ear has a rich population of pericytes, a multi-functional mural cell essential for sensory hair cell heath and normal hearing. However, the mechanics of how pericytes contribute to the homeostasis of the auditory vascular-neuronal complex in the spiral ganglion are not yet known. In this study, using an inducible and conditional pericyte depletion mouse (PDGFRB-CreERT2; ROSA26iDTR) model, we demonstrate, for the first time, that pericyte depletion causes loss of vascular volume and spiral ganglion neurons (SGNs) and adversely affects hearing sensitivity. Using an in vitro trans-well co-culture system, we show pericytes markedly promote neurite and vascular branch growth in neonatal SGN explants and adult SGNs. The pericyte-controlled neural growth is strongly mediated by pericyte-released exosomes containing vascular endothelial growth factor-A (VEGF-A). Treatment of neonatal SGN explants or adult SGNs with pericyte-derived exosomes significantly enhances angiogenesis, SGN survival, and neurite growth, all of which were inhibited by a selective blocker of VEGF receptor 2 (Flk1). Our study demonstrates that pericytes in the adult ear are critical for vascular stability and SGN health. Cross-talk between pericytes and SGNs via exosomes is essential for neuronal and vascular health and normal hearing.

Cell Differentiation-Inspired, Salt-Induced Multifunctional Gels for an Intelligent Soft Robot with an Artificial Reflex Arc

To make soft robotics intelligent, dazzling artificial skin and actuators have been created. However, compared to rigid commercial robots, the sophisticated demands of raw materials become a key challenge for autonomous soft actuators to realize manufacturing repeatability and reproducibility. Inspired by the stem cell, which has the potential to differentiate into multifunctional cells with the same original compositions, a potential multifunctional gel is presented. With well-designed polymer chains, the gel has a salt-induced regulating module, conductivity, and other bionic properties. Making use of this advantage, the gel acts as a double-duty electrode for both the actuator and sensor. An artificial reflex arc is therefore formed by their tight integration via an e-brain: a computing unit that specifically responds to organism intervention. This efficient strategy to obtain diverse components with minimal raw materials is promising for effortlessly fabricating fully soft robotics.

Toolchain for Automated Disassembly for Recycling of Electric Vehicle Batteries

In the context of the increasing focus on the circular economy and the importance of responsible use of resources, more and more attention is being paid to disassembly. While today a large part of the products are shredded and recycled as a single unit, individual components of complex products must be dismantled to reuse or recycle their materials by type. As disassembly is still done manually for many products today, this is only cost-effective for a few. To accelerate the planning process for disassembly and to make automated disassembly using multi-functional robot cells more lucrative, this paper presents a toolchain consisting of a disassembly planning assistant on the one hand and a robot programming assistant on the other. The goal is to simplify and accelerate disassembly planning. As input data, the toolchain receives the product model in a neutral data format and determines disassembly-relevant information from it. A GUI enables the manual adaptation of the data as well as the addition of additional information that cannot be extracted directly from the product model. This information is passed on to a simulation environment, which can be used to intuitively create a template program for a robot in a disassembly cell offline. In addition, evaluation criteria are derived to validate the toolchain.

Construction of a myocardial patch with mesenchymal stem cells and poly(CL-co-TOSUO)/collagen scaffolds for myocardial infarction repair by coaxial electrospinning.

Stem cell cardiac patches have shown promising future for myocardium infarction (MI) therapy, but the characteristics of cardiac pulsation and tissue orientation lead to challenges in the design of cardiac repair scaffolds. Herein, a novel and multifunctional stem cell patch with favorable mechanical properties was reported. In this study, the scaffold was prepared by coaxial electrospinning of poly (CL-co-TOSUO)/collagen (PCT/collagen) core/shell nanofibers. Rat bone marrow-derived mesenchymal stem cells (MSCs) were seeded onto the scaffold to prepare the MSC patch. The nanofiber diameter of coaxial PCT/collagen was 945 ± 102 nm, and the tensile testing results showed that the PCT/collagen core/shell nanofibers possess highly elastic mechanical properties with an elongation at break higher than 300%. The results also showed that the MSCs maintained stem cell properties following seeding on the nano-fibers. The cells on the transplanted MSC patch survived 15 ± 4% 5 weeks following transplantation, and this PCT/collagen-MSC patch significantly improved the MI cardiac function and promoted angiogenesis. With high elasticity and good stem cell biocompatibility, the PCT/collagen core/shell nanofibers exhibited a good research value in the field of myocardial patches.

The role of mast cells in physiological and complicated pregnancy

Mast cells (MCs) are highly differentiated and multifunctional immune cells. The importance of TC has been established not only as mediators of allergic reactions, but also for the development of an immune response, the occurrence of certain autoimmune diseases, tissue homeostasis, the formation of immunotolerance and metastasis of malignant tumors. MCs are present in the endometrium of women in various value depending on age, the phase of the menstrual cycle, the presence of pregnancy. Out of pregnancy, MCs are involved in the cyclic transformation of the uterine mucosa. At the onset of pregnancy, MCs stimulate the process of remodeling of the spiral arteries, the production of leukemia-inhibiting factor (LIF), which is the main implantation factor, and contribute to the formation of an immunotolerant state of the mother in relation to the fetus. Obstetric complications are accompanied by a variable content of MCs, which is associated with different genesis of diseases. A low amount of MCs is associated with impaired implantation and the development of early preeclampsia, an increased content of MCs is observed in the presence of a pathological inflammatory reaction that accompanies late preeclampsia. This review is devoted to the significance of MCs and their mediators in the physiological course of pregnancy, as well as their participation in the pathogenetic mechanisms of pregnancy complications.

Conventional and non-conventional antigen presentation by mast cells.

Mast cells (MCs) are multifunctional immune cells that express a diverse repertoire of surface receptors and pre-stored bioactive mediators. They are traditionally recognized for their involvement in allergic and inflammatory responses, yet there is a growing body of literature highlighting their contributions to mounting adaptive immune responses. In particular, there is growing evidence that MCs can serve as antigen-presenting cells, owing to their often close proximity to T cells in both lymphoid organs and peripheral tissues. Recent studies have provided compelling support for this concept, by demonstrating the presence of antigen processing and presentation machinery in MCs and their ability to engage in classical and non-classical pathways of antigen presentation. However, there remain discrepancies and unresolved questions regarding the extent of the MC's capabilities with respect to antigen presentation. In this review, we discuss our current understanding of the antigen presentation by MCs and its influence on adaptive immunity.

Characteristics of γδTCR on myeloid cells from C57BL/6 mice with Plasmodium yoelii nigeriensis infection

Recently, there is a paucity of studies focus on the characteristics of myeloid cells which expressed γδTCR. The aim of this study was to observe the properties of γδTCR-expressing myeloid cells in the spleen of C57BL/6 mice infected by P. yoelii nigeriensis NSM. Haematoxylin-eosin (HE) staining was used to observe pathological changes in the spleens from infected mice. The differentially expressed genes (DEGs) between the infection and control groups were analyzed by RNA sequencing (RNA -seq). Flow cytometry (FCM) was used to evaluate the frequency of γδTCR+ cells and the characteristics of γδTCR+ cells in P. yoelii nigeriensis NSM-infected mice. Obvious infiltration of inflammatory were observed in the spleens from infected C57BL/6 mouse. The proportions of γδTCR+ cells and CD11b+ γδTCR+ cells from infected group were higher than that from normal group. CD11b+ γδTCR+ cells expressed high levels of activated-mediated genes and inflammatory-mediated genes. The heterogeneous pathway activities among CD11b+ γδTCR+ cells from normal and infected group were characterized. The oxidative phosphorylation, respiratory electron transport chain and leukocyte activation involved in immune response pathways were up-regulated, while the alpha-beta T cell activation and myeloid leukocyte migration pathways were down-regulated in infected mice. Importantly, Ly6c2 was higher expressed in CD11b+ γδTCR+ cells than Ly6g. Consistent with it, flow cytometry results revealed that a subset of Ly6C+ cells was higher than Ly6G+ cells in the spleen. Taken together, our data suggest the existence of a population of γδTCR-expressing myeloid cells and they might be multifunctional cells, which play a role in couse of Plasmodium infection.

Predicted landscape of RETINOBLASTOMA-RELATED LxCxE-mediated interactions across the Chloroplastida.

The colonization of land by a single streptophyte algae lineage some 450million years ago has been linked to multiple key innovations such as three-dimensional growth, alternation of generations, the presence of stomata, as well as innovations inherent to the birth of major plant lineages, such as the origins of vascular tissues, roots, seeds and flowers. Multicellularity, which evolved multiple times in the Chloroplastida coupled with precise spatiotemporal control of proliferation and differentiation were instrumental for theevolution of these traits. RETINOBLASTOMA-RELATED (RBR), the plant homolog of the metazoan Retinoblastoma protein (pRB), is a highly conserved and multifunctional core cell cycle regulator that has been implicated in the evolution of multicellularity in the green lineage as well as in plant multicellularity-related processes such as proliferation, differentiation, stem cell regulation and asymmetric cell division. RBR fulfills these roles through context-specific protein-protein interactions with proteins containing the Leu-x-Cys-x-Glu (LxCxE) short-linear motif (SLiM); however, how RBR-LxCxE interactions have changed throughout major innovations in the Viridiplantae kingdom is a question that remains unexplored. Here, we employ an in silico evo-devo approach to predict and analyze potential RBR-LxCxE interactions in different representative species of key Chloroplastida lineages, providing a valuable resource for deciphering RBR-LxCxE multiple functions. Furthermore, our analyses suggest that RBR-LxCxE interactions are an important component of RBR functions and that interactions with chromatin modifiers/remodelers, DNA replication and repair machinery are highly conserved throughout the Viridiplantae, while LxCxE interactions with transcriptional regulators likely diversified throughout the water-to-land transition.

Assessment of HDAC Inhibitor-Induced Endoplasmic Reticulum (ER) Stress.

The endoplasmic reticulum (ER) is a multifunctional cell organelle which is important for the folding and processing of proteins. Different endogenous and exogenous factors can disturb the ER homeostasis, causing ER stress and activating the unfolded protein response (UPR) to remove misfolded proteins and aggregates. ER stress and the UPR are associated with several human diseases, such as diabetes, Alzheimer's or Parkinson's disease, and cancer. Histone deacetylase inhibitors (HDACi) are used to treat cancer and were shown to induce ER stress/to modulate the UPR, although the exact mechanism is not fully understood and needs further research. Several approaches to monitoring ER stress exist. Here we describe methods including qPCR, Western blot, transmission electron microscopy, and fluorescence microscopy to analyze changes in mRNA and protein expression levels as well as defects in ER structures after HDAC inhibitor-induced ER stress.

Visualizing inflammation with an M1 macrophage selective probe via GLUT1 as the gating target

Macrophages play crucial roles in protecting our bodies from infection and cancers. As macrophages are multi-functional immune cells, they have diverse plastic subsets, such as M1 and M2, derived from naïve M0 cells. Subset-specific macrophage probes are essential for deciphering and monitoring the various activation of macrophages, but developing such probes has been challenging. Here we report a fluorescent probe, CDr17, which is selective for M1 macrophages over M2 or M0. The selective staining mechanism of CDr17 is explicated as Gating-Oriented Live-cell Distinction (GOLD) through overexpressed GLUT1 in M1 macrophages. Finally, we demonstrate the suitability of CDr17 to track M1 macrophages in vivo in a rheumatoid arthritis animal model.

Controversial role of mast cells in NSCLC tumor progression and angiogenesis.

Mast cells (MCs) are multifunctional immune cells implicated in both physiological and pathological processes. Among the latter, MCs play a crucial role in cancer. Many studies have shown a correlation between MCs and tumor progression in several solid and hematological malignancies. In particular, MCs can directly promote tumor growth via c-kit/stem cell factor-dependent signaling and via the release of histamine, which modulate tumor growth through H1 and H2 receptors. At the same time, MCs can increase tumor progression by stimulating angiogenesis via both proangiogenic cytokines stored in their cytoplasm, and by acting on the tumor microenvironment and extracellular matrix. With regard to NSCLC, the role of MCs has not yet been established, with studies showing a correlation with a poor prognosis on the one hand and suggesting a protective effect of MCs on the other hand. These controversial evidences are at least, in part, due to the heterogeneity of the studies exploring the role of MCs in NSCLC, with some studies describing only the MC count without specification of the activation and degranulation state, and without reporting the intratumoral localization and the proximity to other immune and cancer cells. A better knowledge of the role of MCs in NSCLC is mandatory, not only to define their prognostic and predictive proprieties but also because targeting them could be a possible therapeutic strategy.

Oxidized low-density lipoprotein causes ribosome reduction and inhibition of protein synthesis in retinal pigment epithelial cells

Retinal pigment epithelium (RPE) are specialized multifunctional cells indispensable for maintenance of vision. Dysfunction and death of the RPE cells is implicated in the genesis and progression of age-related macular degeneration (AMD). Oxidative stress and resulting cellular damage plays a critical mechanistic role in AMD pathogenesis. Oxidized low-density lipoprotein (oxLDL), derived from LDL in a pro-oxidative environment, is found adjacent to the RPE as part of drusen, extracellular deposits that are a characteristic clinical feature of AMD. OxLDL is cytotoxic and oxLDL-induced oxidative damage may contribute to functional impairment of the RPE. Therefore, knowledge of how the RPE respond to oxLDL exposure is important to understand the mechanisms underlying RPE dysfunction and death associated with AMD. The objective of this study was to characterize alterations in the RPE proteome triggered by exposure to non-cytotoxic levels of oxLDL. Protein identification and quantification were performed with a high -resolution LC-MS/MS-based proteomics workflow. In total, out of the ca 3000 RPE proteins quantified, oxLDL treatment caused expression changes of 303 proteins. As revealed by protein functional analysis, oxLDL uptake caused a multifaceted molecular response that involved numerous biological pathways. This response included up-regulation of anti-oxidative stress proteins whose expression is mediated by the transcription factor nuclear factor erythroid 2-related factor 2 (NRF2), confirming results of transcriptomics studies previously published by us and others. Significantly, and previously unreported, the oxLDL treatment induced down-regulation of ribosomal and translation initiation proteins, and up-regulation of proteins involved in autophagy, thus suggesting that a major cellular mechanism through which the RPE mitigate oxLDL-induced damage involves inhibition of protein synthesis and removal of misfolded proteins.

Dipeptidyl Peptidase-4 Stabilizes Integrin α4β1 Complex to Promote Thyroid Cancer Cell Metastasis by Activating Transforming Growth Factor-Beta Signaling Pathway.

Background: Metastatic disease is a major cause of thyroid cancer-related death. However, the mechanisms responsible for thyroid cancer metastasis are unclear. Dipeptidyl peptidase-4 (DPP4) is a multifunctional cell surface glycoprotein that has been reported to be a negative prognostic factor in thyroid cancer. We explored the molecular mechanism of the role of DPP4 in thyroid cancer cell metastasis. Methods: The effects of DPP4 on thyroid cancer cell migration/invasion in vitro were assessed by transwell assays. A lung metastatic mouse model was also established to determine the effect of DPP4 on tumor metastasis in vivo. DPP4 inhibitor sitagliptin was used to test its effect on thyroid cancer cell metastasis. The mechanism of which DPP4 promotes thyroid cancer cell metastasis was explored by a series of molecular and biochemical experiments. Results: We observed that DPP4 was significantly upregulated in papillary thyroid cancers compared with control subjects, and its expression was positively associated with lymph node metastasis and BRAFV600E mutation. Functional studies showed that DPP4 knockdown significantly inhibited metastatic potential of thyroid cancer cells, and vice versa. However, DPP4 inhibitor sitagliptin did not affect the metastatic ability of thyroid cancer cells, indicating that the promoting effect of DPP4 on tumor metastasis was independent of its enzymatic activity. Mechanistically, DPP4 interacted with the α4 and β1 integrin subunits, and stabilized the formation of integrin α4β1 complex. DPP4-mediated integrin signal activation promoted the nuclear localization of c-Jun through the FAK/AKT pathway, thereby inducing the transcription of transforming growth factor-beta 1 (TGFB1 coding for protein TGF-β1). TGF-β1 then facilitated tumor metastasis by inducing the epithelial-mesenchymal transition. Conclusions: DPP4 promotes thyroid cancer cell metastasis through the integrins/FAK/AKT/c-Jun/TGF-β1 signaling axis. These findings may have implications for an alternative therapeutic strategy for thyroid cancer.

3차원 구조를 갖는 유연 및 신축 소자

Developments of ways to fabricate complex three-dimensional (3D) structures enable the controlling of physical and chemical properties of the electrical systems. Especially, owing to the rapid developments of the fabrication processes (e.g., the 3D printing, origami, and mechanical buckling process), researchers have integrated multifunctional and sophisticated 3D structures with flexible and stretchable substrates for high electrical, mechanical, and optical performances. In this review, we highlight the latest research on flexible and stretchable electric systems integrated with 3D structures such as a super-sensitive pressure sensor, a high-performance wearable monitoring system, a multi-functional cell scaffold, a foldable thermoelectric generator, a wearable energy harvesting system, a hemispherical photodetector array, and a projection screen with the reversible state changes. In subsequent sections, we summarize the advanced research results and provide future strategies for flexible and stretchable 3D electronic devices.

CD95 gene deletion may reduce clonogenic growth and invasiveness of human glioblastoma cells in a CD95 ligand-independent manner

CD95 (Fas/APO-1) is a multifunctional cell surface receptor with antithetic roles. First described to mediate cell death, interactions of CD95 with its natural ligand, CD95L, have also been described to induce tumor-promoting signaling leading to proliferation, invasion and stem cell maintenance, mainly in cancer cells that are resistant to CD95-mediated apoptosis. While activation of CD95-mediated apoptosis in cancer cells may not be clinically practicable due to toxicity, inhibition of tumor-promoting CD95 signaling holds therapeutic potential. In the present study, we characterized CD95 and CD95L expression in human glioma-initiating cells (GIC), a glioblastoma cell population with stem cell features, and investigated the consequences of CRISPR-Cas9-mediated CD95 or CD95L gene deletion. In vitro, GIC expressed CD95 but not CD95L and were sensitive to CD95-mediated apoptosis. Upon genetic deletion of CD95, GIC acquired resistance to CD95L-induced apoptosis but exhibited inferior clonogenic growth, sphere-forming capacity, and invasiveness compared with control cells, suggesting the existence of CD95L-independent constitutive CD95 signaling with tumor-promoting properties in GIC. In vivo, GIC expressed CD95 and a non-canonical form of CD95L lacking the CD95-binding region. CD95 genetic deletion did not prolong survival in immunocompromised GIC-bearing mice. Altogether, these data indicate that canonical CD95L may not be expressed in human GIC and suggest the existence of a CD95L-independent CD95-signaling pathway that maintains some malignancy traits of GIC. The lack of altered survival of tumor-bearing mice after genetic deletion of CD95 suggests that CD95 signaling is not essential to maintain the growth of human GIC xenografted into the brains of nude mice. The ligand-independent tumor-promoting role of constitutive CD95 in our GIC models in vitro highlights the complexity and challenges associated with targeting CD95 with therapeutic intent.

Autoantibodies to IgE can induce the release of proinflammatory and vasoactive mediators from human cardiac mast cells

Mast cells are multifunctional immune cells with complex roles in tissue homeostasis and disease. Cardiac mast cells (HCMCs) are strategically located within the human myocardium, in atherosclerotic plaques, in proximity to nerves, and in the aortic valve. HCMCs express the high-affinity receptor (FcεRI) for IgE and can be activated by anti-IgE and anti-FcεRI. Autoantibodies to IgE and/or FcεRI have been found in the serum of patients with a variety of immune disorders. We have compared the effects of different preparations of IgG anti-IgE obtained from patients with atopic dermatitis (AD) with rabbit IgG anti-IgE on the release of preformed (histamine and tryptase) and lipid mediators [prostaglandin D2 (PGD2) and cysteinyl leukotriene C4 (LTC4)] from HCMCs. Functional human IgG anti-IgE from one out of six AD donors and rabbit IgG anti-IgE induced the release of preformed (histamine, tryptase) and de novo synthesized mediators (PGD2 and LTC4) from HCMCs. Human IgG anti-IgE was more potent than rabbit IgG anti-IgE in inducing proinflammatory mediators from HCMCs. Human monoclonal IgE was a competitive antagonist of both human and rabbit IgG anti-IgE. Although functional anti-IgE autoantibodies rarely occur in patients with AD, when present, they can powerfully activate the release of proinflammatory and vasoactive mediators from HCMCs.

A hybrid shape memory polymer filled metallic foam composite: shape restoring, strain sensing, Joule heating, strengthening, and toughening

In this paper, an open-cell metallic foam was filled in by a tough shape memory polymer (SMP), to form a hybrid metal/polymer composite with multifunctionalities and enhanced mechanical properties. This work aims to study the positive composite actions between the metallic skeleton and the SMP filler. Mechanical, thermal, and conductive properties of the resulting hybrid composite were evaluated and compared to the individual components. Uniaxial compression tests and shape memory effect tests were conducted. Results demonstrated an improvement in the compressive strength and toughness. The hybrid composite also exhibited excellent shape recovery and high recovery stress of 1.76 MPa. Infrared thermography has been used to verify the free shape recovery by Joule heating. Sandwich structures with the hybrid composite as the core were studied through low velocity impact test and three-point bending test. The sandwich structures with the composite foam core showed significant performance improvement in both tests. Electrical resistivity study during the three-point bending test validates the possible application of this multifunctional polymer-aluminum open cell foam composite as strain sensor. This type of hybrid composites can be beneficial in many industrial sectors that search for an ideal combination of high strength, high toughness, low weight, damage sensing, and excellent energy absorption capabilities.

Development of myelinating glia: An overview.

Myelin is essential to nervous system function, playing roles in saltatory conduction and trophic support. Oligodendrocytes (OLs) and Schwann cells (SCs) form myelin in the central and peripheral nervous systems respectively and follow different developmental paths. OLs are neural stem-cell derived and follow an intrinsic developmental program resulting in a largely irreversible differentiation state. During embryonic development, OL precursor cells (OPCs) are produced in distinct waves originating from different locations in the central nervous system, with a subset developing into myelinating OLs. OPCs remain evenly distributed throughout life, providing a population of responsive, multifunctional cells with the capacity to remyelinate after injury. SCs derive from the neural crest, are highly dependent on extrinsic signals, and have plastic differentiation states. SC precursors (SCPs) are produced in early embryonic nerve structures and differentiate into multipotent immature SCs (iSCs), which initiate radial sorting and differentiate into myelinating and non-myelinating SCs. Differentiated SCs retain the capacity to radically change phenotypes in response to external signals, including becoming repair SCs, which drive peripheral regeneration. While several transcription factors and myelin components are common between OLs and SCs, their differentiation mechanisms are highly distinct, owing to their unique lineages and their respective environments. In addition, both OLs and SCs respond to neuronal activity and regulate nervous system output in reciprocal manners, possibly through different pathways. Here, we outline their basic developmental programs, mechanisms regulating their differentiation, and recent advances in the field.

Abstract 4233: Generation of a Bicycle NK-TICA™, a novel NK cell engaging molecule designed to induce targeted tumor cytotoxicity

Abstract The tumor specific activation of natural killer (NK) cells is an area of active investigation in immune oncology, but to date has relied on complex biologic modalities (e.g., antibodies, fusion proteins, or cell therapies). NK cells are highly responsive immune cells that can detect and eliminate tumor cells and bridge innate to adaptive immune responses. Bicycles® are small (ca.1.5kDa), chemically synthetic, structurally constrained peptides discovered via phage display and optimized using structure-driven design and medicinal chemistry approaches. We have applied the Bicycle platform technology to identify Bicycles® that bind specifically to the key activating receptor, NKp46. When chemically coupled to tumor antigen binding Bicycles, this results in highly potent, antigen-dependent receptor activation and NK cell activation. We term this new class of fully synthetic molecules NK-TICAs and we will describe herein their discovery and evaluation.We demonstrate potent, selective binding of our Bicycles to receptor-expressing cells and the capability of the bifunctional molecule to induce NK cell function in vitro. With Bicycle’s novel NK-TICA™ compound, we demonstrate the engagement of NK cells, the specific activation and function of NK cells, and enhanced tumor cytotoxicity in a tumor target- and dose-dependent manner.In conclusion, NK-TICAs drive NK cell-mediated tumor cell killing and cytokine production in vitro and as such have the potential to catalyze the development of durable anti-tumor immunity in tumor types not well served by current therapies. We hypothesize that utilization of Bicycle NK-TICA™ as a multifunctional immune cell engager will promote the modulation of NK cells, as well as the infiltration and anti-tumor activity of NK cells in solid tumors. The data presented here provide initial proof of concept for the application of our Bicycle technology to drive NK cell-mediated tumor immunity. Citation Format: Fay J. Dufort, Christopher J. Leitheiser, Gemma Mudd, Julia Kristensson, Alexandra Rezvaya, Katie Gaynor, Sandra Uhlenbroich, Liudvikas Urbonas, Heather Scott, Liuhong Chen, Helen Harrison, Michael Skynner, Kevin McDonnell, Philip E. Brandish, Nicholas Keen. Generation of a Bicycle NK-TICA™, a novel NK cell engaging molecule designed to induce targeted tumor cytotoxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 4233.