RESUMO
Improving drug delivery efficiency to solid tumor sites is a central challenge in anticancer therapeutic research. Our previous experimental study (Guo et al., Nat. Commun. 2018, 9, 130) showed that soft, elastic liposomes had increased uptake and accumulation in cancer cells and tumors in vitro and in vivo respectively, relative to rigid particles. As a first step toward understanding how liposomes' molecular structure and composition modulates their elasticity, we performed all-atom and coarse-grained classical molecular dynamics (MD) simulations of lipid bilayers formed by mixing a long-tailed unsaturated phospholipid with a short-tailed saturated lipid with the same headgroup. The former types of phospholipids considered were 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine (termed here DPMPC). The shorter saturated lipids examined were 1,2-diheptanoyl-sn-glycero-3-phosphocholine (DHPC), 1,2-didecanoyl-sn-glycero-3-phosphocholine (DDPC), 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). Several lipid concentrations and surface tensions were considered. Our results show that DOPC or DPMPC systems having 25-35 mol % of the shortest lipids DHPC or DDPC are the least rigid, having area compressibility moduli KA that are â¼10% smaller than the values observed in pure DOPC or DPMPC bilayers. These results agree with experimental measurements of the stretching modulus and lysis tension in liposomes with the same compositions. These mixed systems also have lower areas per lipid and form more uneven x-y interfaces with water, the tails of both primary and secondary lipids are more disordered, and the terminal methyl groups in the tails of the long lipid DOPC or DPMPC wriggle more in the vertical direction, compared to pure DOPC or DPMPC bilayers or their mixtures with the longer saturated lipid DLPC or DMPC. These observations confirm our hypothesis that adding increasing concentrations of the short unsaturated lipid DHPC or DDPC to DOPC or DPMPC bilayers alters lipid packing and thus makes the resulting liposomes more elastic and less rigid. No formation of lipid nanodomains was noted in our simulations, and no clear trends were observed in the lateral diffusivities of the lipids as the concentration, type of secondary lipid, and surface tension were varied.
Assuntos
Lipossomos , Simulação de Dinâmica Molecular , Lipossomos/química , Dimiristoilfosfatidilcolina/química , Fosforilcolina , Fosfolipídeos/química , Bicamadas Lipídicas/química , Fosfatidilcolinas/químicaRESUMO
Lysophosphatidic acid receptor 1 (LPAR1) is elevated in breast cancer. The deregulation of LPAR1, including the function and level of expression, is linked to cancer initiation, progression, and metastasis. LPAR1 antagonists, AM095 or Ki16425, may be effective therapeutic molecules, yet their limited water solubility hinders in vivo delivery. In this study, we report on the synthesis of two liposomal formulations incorporating AM095 or Ki16425, embedded within the lipid bilayer, as targeted nanocarriers for metastatic breast cancer (MBC). The data show that the Ki16425 liposomal formulation exhibited a 50% increase in internalization by MBC mouse epithelial cells (4T1) and a 100% increase in tumor accumulation in a mouse model of MBC compared with that of a blank liposomal formulation (control). At the same time, normal mouse epithelial cells (EpH-4Ev) internalized the Ki16425 liposomal formulation 25% lesser than the control formulation. Molecular dynamics simulations show that the integration of AM095 or Ki16425 modified the physical and mechanical properties of the lipid bilayer, making it more flexible in these liposomal formulations compared with liposomes without drug. The incorporation of an LPAR1 antagonist within a liposomal drug delivery system represents a viable therapeutic approach for targeting the LPA-LPAR1 axis, which may hinder the progression of MBC.
Assuntos
Neoplasias da Mama , Lipossomos , Humanos , Camundongos , Animais , Feminino , Neoplasias da Mama/tratamento farmacológico , Bicamadas Lipídicas , Modelos Animais de Doenças , Receptores de Ácidos Lisofosfatídicos/metabolismoRESUMO
Liposomal drug delivery represents a highly adaptable therapeutic platform for treating a wide range of diseases. Natural and synthetic lipids, as well as surfactants, are commonly utilized in the synthesis of liposomal drug delivery vehicles. The molecular diversity in the composition of liposomes enables drug delivery with unique physiological functions, such as pH response, prolonged blood circulation, and reduced systemic toxicity. Herein, we discuss the impact of composition on liposome synthesis, function, and clinical utility.
Assuntos
Sistemas de Liberação de Medicamentos , Desenho de Fármacos , Lipídeos/química , Animais , Humanos , Concentração de Íons de Hidrogênio , Lipossomos , Preparações Farmacêuticas/administração & dosagem , Preparações Farmacêuticas/química , Tensoativos/químicaRESUMO
Triple-negative breast cancer (TNBC), which has the highest mortality rate of all breast cancer, is in urgent need of a therapeutic that hinders the spread and growth of cancer cells. CRISPR genome editing holds the promise of a potential cure for many genetic diseases, including TNBC; however, its clinical translation is being challenged by the lack of safe and effective nonviral delivery systems for in vivo therapeutic genome editing. Here we report the synthesis and application of a noncationic, deformable, and tumor-targeted nanolipogel system (tNLG) for CRISPR genome editing in TNBC tumors. We have demonstrated that tNLGs mediate a potent CRISPR knockout of Lipocalin 2 (Lcn2), a known breast cancer oncogene, in human TNBC cells in vitro and in vivo. The loss of Lcn2 significantly inhibits the migration and the mesenchymal phenotype of human TNBC cells and subsequently attenuates TNBC aggressiveness. In an orthotopic TNBC model, we have shown that systemically administered tNLGs mediated >81% CRISPR knockout of Lcn2 in TNBC tumor tissues, resulting in significant tumor growth suppression (>77%). Our proof-of-principle results provide experimental evidence that tNLGs can be used as a safe, precise, and effective delivery approach for in vivo CRISPR genome editing in TNBC.
Assuntos
Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Edição de Genes , Nanopartículas/química , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/genética , Animais , Sistemas CRISPR-Cas , Linhagem Celular Tumoral , Movimento Celular , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Terapia Genética/métodos , Vetores Genéticos , Humanos , Molécula 1 de Adesão Intercelular/genética , Lipídeos/química , Lipocalina-2/genética , Lipossomos/química , Camundongos , Camundongos NusRESUMO
The use of hydrogels in load bearing applications is often limited by insufficient toughness. 2-Hydroxyethyl methacrylate (HEMA) based hydrogels are appealing for translational work, as they are affordable and the use of HEMA is FDA approved. Furthermore, HEMA is photopolymerizable, providing spatiotemporal control over mechanical properties. We evaluated the ability of vinyl methacrylate (VM), allyl methacrylate (AM), and 3-(Acryloyloxy)-2-hydroxypropyl methacrylate (AHPM) to tune hydrogel toughness and Young's modulus. The crosslinkers were selected due to their heterobifunctionality (vinyl and methacrylate) and similar size and structure to EGDMA, which was shown previously to increase toughness as compared to longer crosslinkers. Vinyl methacrylate incorporation into HEMA hydrogels gave rise to hydrogels with Young's moduli spanning ranges for ligament to cartilage, with a peak toughness of 519 ± 70 kJ/m3 under physiological conditions. We report toughness (work of extension) as a function of modulus and equilibrium water content for all formulations. The hydrogels exhibited 80%-100% cell viability, which suggests they could be used in tissue engineering applications.
Assuntos
Materiais Biocompatíveis/química , Hidrogéis/química , Fenômenos Mecânicos , Metacrilatos/química , Materiais Biocompatíveis/farmacologia , Cartilagem/citologia , Cartilagem/efeitos dos fármacos , Humanos , Teste de Materiais , Metacrilatos/farmacologia , Processos Fotoquímicos , Polimerização , Água/químicaRESUMO
The plasticity of cancer epigenetics makes them plausible candidates for therapeutic intervention. We took advantage of elevated expression of lysophosphatidic acid receptor 1 (LPAR1) in triple negative breast cancer (TNBC) tissues to target decitabine (DAC) and panobinostat (PAN) to breast cancer cells. DAC and PAN were shown to reverse abnormal methylation of DNA and altered chromatin structure, respectively, leading to increased expression of tumor suppressor genes and decreased expression of oncogenes. Although DAC and PAN have therapeutic benefits, they are limited by chemical instability and systemic toxicity. Herein, we present LPAR1-targeted, lipid nanoemulsions (LNEs) encapsulating both DAC and PAN. Our results demonstrated that the cell uptake and in vivo biodistribution of LNEs was dependent on LPAR1 expression in TNBCs. DAC/PAN-LNEs were effective in inhibiting the growth of mesenchymal breast cancer cells by restoring CDH1/E-cadherin and suppressing forkhead box M1 (FOXM1) expression. Epithelial breast cancer cells that inherently express low FOXM1 and high CDH1 were unaffected by DAC/PAN-LNEs. Overall, we successfully designed LPAR1-targeted LNEs that selectively act on CDH1(low)/FOXM1(high) TNBC cell lines.
Assuntos
Antígenos CD/metabolismo , Antimetabólitos Antineoplásicos/farmacocinética , Caderinas/metabolismo , Decitabina/farmacocinética , Proteína Forkhead Box M1/metabolismo , Lipídeos/química , Nanocápsulas/química , Panobinostat/farmacocinética , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Antimetabólitos Antineoplásicos/uso terapêutico , Proliferação de Células/efeitos dos fármacos , Decitabina/uso terapêutico , Desenho de Fármacos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Feminino , Xenoenxertos , Células Endoteliais da Veia Umbilical Humana , Humanos , Células MCF-7 , Camundongos , Camundongos Nus , Panobinostat/uso terapêutico , Receptores de Ácidos Lisofosfatídicos/metabolismo , Transdução de Sinais/efeitos dos fármacos , Distribuição Tecidual , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
Distinguishing malignant cells from non-neoplastic ones is a major challenge in triple-negative breast cancer (TNBC) treatment. Here, we developed a complementary targeting strategy that uses precisely matched, multivalent ligand-receptor interactions to recognize and target TNBC tumors at the primary site and metastatic lesions. We screened a panel of cancer cell surface markers and identified intercellular adhesion molecule-1 (ICAM1) and epithelial growth factor receptor (EGFR) as optimal candidates for TNBC complementary targeting. We engineered a dual complementary liposome (DCL) that precisely complements the molecular ratio and organization of ICAM1 and EGFR specific to TNBC cell surfaces. Our in vitro mechanistic studies demonstrated that DCLs, compared to single-targeting liposomes, exhibited increased binding, enhanced internalization, and decreased receptor signaling. DCLs consistently exhibited substantially increased tumor targeting activity and antitumor efficacy in orthotopic and lung metastasis models, indicating that DCLs are a platform technology for the design of personalized nanomedicines for TNBC.
Assuntos
Molécula 1 de Adesão Intercelular/genética , Lipossomos/farmacologia , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Progressão da Doença , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Feminino , Humanos , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Lipossomos/metabolismo , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Camundongos , Terapia de Alvo Molecular , Metástase Neoplásica , Transdução de Sinais , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Receptor-mediated drug delivery presents an opportunity to enhance therapeutic efficiency by accumulating drug within the tissue of interest and reducing undesired, off-target effects. In cancer, receptor overexpression is a platform for binding and inhibiting pathways that shape biodistribution, toxicity, cell binding and uptake, and therapeutic function. This review will identify tumor-targeted drug delivery vehicles and receptors that show promise for clinical translation based on quantitative in vitro and in vivo data. The authors describe the rationale to engineer a targeted drug delivery vehicle based on the ligand, chemical conjugation method, and type of drug delivery vehicle. Recent advances in multivalent targeting and ligand organization on tumor accumulation are discussed. Revolutionizing receptor-mediated drug delivery may be leveraged in the therapeutic delivery of chemotherapy, gene editing tools, and epigenetic drugs.
RESUMO
To date, the role of elasticity in drug delivery remains elusive due to the inability to measure microscale mechanics and alter rheology without affecting chemistry. Herein, we describe the in vitro cellular uptake and in vivo tumor uptake of nanolipogels (NLGs). NLGs are composed of identical lipid bilayers encapsulating an alginate core, with tunable elasticity. The elasticity of NLGs was evaluated by atomic force microscopy, which demonstrated that they exhibit Young's moduli ranging from 45 ± 9 to 19,000 ± 5 kPa. Neoplastic and non-neoplastic cells exhibited significantly greater uptake of soft NLGs (Young's modulus <1.6 MPa) relative to their elastic counterparts (Young's modulus >13.8 MPa). In an orthotopic breast tumor model, soft NLGs accumulated significantly more in tumors, whereas elastic NLGs preferentially accumulated in the liver. Our findings demonstrate that particle elasticity directs tumor accumulation, suggesting that it may be a design parameter to enhance tumor delivery efficiency.
Assuntos
Neoplasias da Mama/metabolismo , Módulo de Elasticidade , Nanopartículas/química , Nanopartículas/metabolismo , Animais , Neoplasias da Mama/patologia , Linhagem Celular , Linhagem Celular Tumoral , Clorpromazina/farmacologia , Endocitose/efeitos dos fármacos , Filipina/farmacologia , Humanos , Hidrazonas/farmacologia , Fígado/metabolismo , Células MCF-7 , Neoplasias Mamárias Experimentais/metabolismo , Camundongos Endogâmicos BALB C , Microscopia de Força AtômicaRESUMO
Trachea replacement for nonoperable defects remains an unsolved problem due to complications with stenosis and mechanical insufficiency. While native trachea has anisotropic mechanical properties, the vast majority of engineered constructs focus on uniform cartilaginous-like conduits. These conduits often lack quantitative mechanical analysis at the construct level, which limits analysis of functional outcomes in vivo, as well as comparisons across studies. This review aims to present a clear picture of native tracheal mechanics at the tissue and organ level, as well as loading conditions to establish design criteria for trachea replacements. We further explore the implications of failing to match native properties with regards to implant collapse, stenosis, and infection.
RESUMO
Breast cancer is the second leading cause of cancer-related mortality in women. Successful development of sensitive nanoprobes for breast cancer cell detection is of great importance for breast cancer diagnosis and symptomatic treatment. Herein, inspired by the intrinsic peroxidase property of gold nanoclusters, high loading, and targeting ability of ErbB2/Her2 antibody functionalized liposomes, we report that gold nanoclusters-loaded, target-directed, functionalized liposomes can serve as a robust sensing platform for amplified colorimetric detection of HER2-positive breast cancer cells. This approach allows HER2-positive breast cancer cell identification at high sensitivity with high selectivity. In addition, the colorimetric "readout" offers extra advantages in terms of low-cost, portability, and easy-to-use applications. The practicality of this platform was further proved by successful detection of HER2-positive breast cancer cells in human serum samples and in breast cancer tissue, which indicated our proposed method has potential for application in cancer theranostics.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Colorimetria/métodos , Ouro/metabolismo , Lipossomos/metabolismo , Técnicas de Diagnóstico Molecular/métodos , Nanopartículas/metabolismo , Receptor ErbB-2/análise , Anticorpos/imunologia , Humanos , Receptor ErbB-2/imunologia , Sensibilidade e EspecificidadeRESUMO
Identifying a molecular target is essential for tumor-targeted nanomedicine. Current cancer nanomedicines commonly suffer from poor tumor specificity, "off-target" toxicity, and limited clinical efficacy. Here, we report a method to screen and identify new molecular targets for tumor-targeted nanomedicine based on a quantitative analysis. In our proof-of-principle study, we used comparative flow cytometric screening to identify ICAM-1 as a potential target for metastatic melanoma (MM). We further evaluated ICAM-1 as a MM targeting moiety by characterizing its (1) tumor specificity, (2) expression level, (3) cellular internalization, (4) therapeutic function, and (5) potential clinical impact. Quantitation of ICAM-1 protein expression on cells and validation by immunohistochemistry on human tissue specimens justified the synthesis of antibody-functionalized drug delivery vehicles, which were benchmarked against appropriate controls. We engineered ICAM-1 antibody conjugated, doxorubicin encapsulating immunoliposomes (ICAM-Dox-LPs) to selectively recognize and deliver doxorubicin to MM cells and simultaneously neutralize ICAM-1 signaling via an antibody blockade, demonstrating significant and simultaneous inhibitory effects on MM cell proliferation and migration. This paper describes a novel, quantitative metric system that identifies and evaluates new cancer targets for tumor-targeting nanomedicine.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Antineoplásicos/administração & dosagem , Doxorrubicina/análogos & derivados , Molécula 1 de Adesão Intercelular/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Criança , Doxorrubicina/administração & dosagem , Feminino , Citometria de Fluxo , Humanos , Molécula 1 de Adesão Intercelular/genética , Masculino , Melanócitos , Melanoma/genética , Melanoma/metabolismo , Pessoa de Meia-Idade , Terapia de Alvo Molecular , Nanomedicina , Polietilenoglicóis/administração & dosagem , Adulto JovemRESUMO
There remains no routine treatment for congenital tracheal abnormalities affecting more than 1/3 of the length. Natural and artificial prostheses are plagued by mechanical failure and inconsistent outcomes. Mimicking native tissue mechanics in an engineered replacement may improve functional and patient outcomes. We synthesized tubular constructs comprising photo-cross-linked methyl acrylate-co-methyl methacrylate, p(MA-co-MMA), with patterned r- and z-axes in order to achieve mechanical properties similar to lamb tracheae. Hard and soft alternating bands, and a soft vertical section, mimic tracheal architecture. Patterned constructs were capable of 46% elastic longitudinal extension. The construct longitudinal composite modulus, 0.34 ± 0.09 MPa, was not significantly different from ovine tracheae. The superior of two geometries evaluated supports up to a 46% reduction of internal volume within the physiological range of transmural pressures. Thus, these patterned hydrogels yielded longitudinal elasticity and radial rigidity while allowing for radial deformation required for effective coughing.
RESUMO
Early detection of breast cancer is a critical component in patient prognosis and establishing effective therapy regimens. Here, we developed an easily accessible yet potentially powerful sensor to detect cancer cell targets by utilizing seven dual-ligand cofunctionalized gold nanoclusters (AuNCs) as both effective cell recognition elements and signal transducers. On the basis of this AuNC multichannel sensor, we have successfully distinguished healthy, cancerous and metastatic human breast cells with excellent reproducibility and high sensitivity. Triple negative breast cancer cells (TNBCs), which exhibit low expression of the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor-2, were identified. The high accuracy of the blind breast cell sample tests further validates the practical application of the sensor array. In addition, the versatility of the sensor array is further justified by identifying amongst distinct cell types, different cell concentrations and cell mixtures. Notably, the drug-resistant cancer cells can also be efficiently discriminated. Furthermore, the dual-ligand cofunctionalized AuNCs can efficiently differentiate different cells from the peripheral blood of tumor-free and tumor-bearing mice. Taken together, this fluorescent AuNCs based array provides a powerful cell analysis tool with potential applications in biomedical diagnostics.
Assuntos
Biomarcadores Tumorais/análise , Ouro/química , Nanopartículas Metálicas/química , Nanoconjugados/química , Espectrometria de Fluorescência , Neoplasias de Mama Triplo Negativas/química , Neoplasias de Mama Triplo Negativas/diagnóstico , Algoritmos , Linhagem Celular Tumoral , Humanos , Ligantes , Reconhecimento Automatizado de Padrão , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The linker between the targeting moiety and the nanoparticle is often overlooked when engineering targeted drug delivery vehicles. We hypothesized that pH-triggered conformational changes of an elastin-like peptide (ELP) linker, with repeating VPGVG sequences, could alter the binding affinity of the well-established targeting moiety arginine-glycine-aspartic acid (RGD), which is known to enhance the delivery of nanoparticles to tumor cells via integrin overexpression. The pH change from blood (pH 7.4) to the tumor environment (pH 6) was used to elicit a conformational change in the ELP linker, as described by circular dichroism. Atomic force microscopy confirmed that RGD-ELP resulted in stronger adhesion to both MDA-MB-231 and HCC1806 breast cancer cells at pH 6 relative to pH 7.4. No change in adhesion force was measured as a function of pH for the non-neoplastic MCF-10A cell line and the nontargeting GDR-ELP peptide. This translated to significant binding and uptake of RGD-ELP modified liposomes at pH 6.0 relative to pH 7.4. These results indicate that the pH-triggered conformational structure of the ELP linker shifts RGD-mediated cancer cell targeting from non-active (pH 7.4) to active (pH 6). The reversible shift in ELP secondary structure may be used to engineer targeted drug delivery vehicles with tunable uptake.
Assuntos
Elastina/química , Oligopeptídeos/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Sequência de Aminoácidos , Adesão Celular , Linhagem Celular Tumoral , Humanos , Concentração de Íons de Hidrogênio , Integrina alfa5beta1/química , Integrina alfa5beta1/metabolismo , Integrina alfaVbeta3/química , Integrina alfaVbeta3/metabolismo , Lipossomos , Modelos Moleculares , Nanopartículas/química , Fragmentos de Peptídeos/toxicidade , Estrutura Secundária de Proteína , Transporte ProteicoRESUMO
Early and accurate diagnosis of breast cancer holds great promise to improve treatability and curability. Here, we report the usage of six luminescent nanodot-graphene oxide complexes as novel fluorescent nanoprobes in a sensing array capable of effectively identifying healthy, cancerous, and metastatic human breast cells. The sensory system is based on the utilization of nanoprobe-graphene oxide sensor elements that can be disrupted in the presence of breast cells to give fluorescent readouts. Using this multichannel sensor, we have successfully identified breast cancer cells and distinguished between estrogen receptor positive, human epidermal growth factor receptor-2 positive, and triple negative phenotypes. This approach also allows cell identification at high sensitivity (200 cells) with high reproducibility. The unknown cell sample analysis indicates that the sensor is able to identify 49 out of 50 breast cell samples correctly, with a detection accuracy of 98%. Taken together, this array-based luminescent nanoprobe-graphene oxide sensing platform presents a useful cell screening tool with potential applications in biomedical diagnostics.
Assuntos
Técnicas Biossensoriais/métodos , Mama/patologia , Corantes Fluorescentes/química , Grafite/química , Nanopartículas/química , Óxidos/química , Neoplasias de Mama Triplo Negativas/diagnóstico , Linhagem Celular Tumoral , Feminino , Humanos , Nanopartículas/ultraestrutura , Reprodutibilidade dos Testes , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
Lipocalin 2 (Lcn2) is a promising therapeutic target as well as a potential diagnostic biomarker for breast cancer. It has been previously shown to promote breast cancer progression by inducing the epithelial to mesenchymal transition in breast cancer cells as well as by enhancing angiogenesis. Lcn2 levels in urine and tissue samples of breast cancer patients has also been correlated with breast cancer status and poor patient prognosis. In this study, we have engineered a novel liposomal small interfering RNA (siRNA) delivery system to target triple negative breast cancer (TNBC) via a recently identified molecular target, intercellular adhesion molecule-1 (ICAM-1). This ICAM-1-targeted, Lcn2 siRNA- encapsulating liposome (ICAM-Lcn2-LP) binds human TNBC MDA-MB-231cells significantly stronger than non-neoplastic MCF-10A cells. Efficient Lcn2 knockdown by ICAM-Lcn2-LPs led to a significant reduction in the production of vascular endothelial growth factor (VEGF) from MDA-MB-231 cells, which, in turn, led to reduced angiogenesis both in vitro and in vivo. Angiogenesis (neovascularization) is a requirement for solid tumor growth and progression, and its inhibition is an important therapeutic strategy for human cancers. Our results indicate that a tumor-specific strategy such as the TNBC-targeted, anti-angiogenic therapeutic approach developed here, may be clinically useful in inhibiting TNBC progression.
Assuntos
Proteínas de Fase Aguda/antagonistas & inibidores , Inibidores da Angiogênese/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Lipocalinas/antagonistas & inibidores , Lipossomos/metabolismo , Terapia de Alvo Molecular/métodos , Proteínas Proto-Oncogênicas/antagonistas & inibidores , RNA Interferente Pequeno/metabolismo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/fisiologia , Humanos , Vírus da Leucemia Murina , Lipocalina-2 , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Tracheal damage, abnormality or absence can result from the growth of tumors or from Congenital High Airway Obstruction Syndrome. No optimal or routine treatment has been established for tracheal repair, despite numerous attempts with natural and artificial prostheses. The fetal trachea is comprised of cartilaginous rings connected by an elastomeric tissue. In an effort to design an engineered trachea replacement, we have synthesized 2-hydroxyethyl methacrylate hydrogels with moduli of 67 ± 3.1 kPa (soft) and 13.0 ± 1.8 MPa (hard). Given the criteria for longitudinal extensibility and lateral rigidity applied during respiration, we evaluated a series of patterned hydrogels with different sizes of hard and soft segments to mimic fetal tracheas. A 1:2 ratio of soft:hard segments resulted in a construct capable of 11.0 ± 1% extension within the elastic range. Tubular constructs with this ratio required similar load/length for cyclic compression as ovine trachea samples. Achieving biomimetic mechanical properties in a trachea replacement may be essential for achieving normal respiration in recipient patients. STATEMENT OF SIGNIFICANCE: Fetal abnormalities or tumors can result in tracheal absence or damage. Despite numerous attempts with natural and artificial replacements, there is still no routine treatment for tracheal repair. The literature recognizes the importance of tracheal lateral rigidity and longitudinal extensibility for normal respiration. Achieving closely matched mechanical properties may provide proper function and help decrease implant fibrosis and subsequent occlusion. In this study, we evaluated the mechanics of a series of patterned, tubular hydrogels with different ratios of hard and soft segments to mimic alternating cartilage and ligament sections in fetal tracheas. We compared our results to that of sheep trachea. This is the first report to assess both radial rigidity and longitudinal extensibility in an engineered trachea construct.
Assuntos
Alicerces Teciduais/química , Traqueia/fisiologia , Animais , Animais Recém-Nascidos , Fenômenos Biomecânicos , Força Compressiva , Módulo de Elasticidade , Técnicas In Vitro , Ovinos , TemperaturaRESUMO
Leukocyte recruitment plays a key role in chronic inflammatory diseases such as cardiovascular disease, rheumatoid arthritis, and cancer. Leukocyte rolling and arrest are mediated in part by the temporally-regulated surface expression of vascular cell adhesion molecule-1 (VCAM1) on endothelial cells (ECs). In this paper, we engineered a pH-responsive vehicle comprised of 30 mol% dimethylaminoethyl methacrylate (30D) and 70 mol% hydroxyethyl methacrylate (70H) to encapsulate, protect, and deliver VCAM1 small interfering RNA (siRNA). The ability of siRNA to reduce VCAM1 gene expression is in direct opposition to its activation by cytokines. At 12 h post-activation, VCAM1 gene knockdown was 90.1 ± 7.5% when delivered via 30D/70H nanoparticles, which was on par with a leading commercial transfection agent. This translated into a 68.8 ± 6.7% reduction in the surface density of VCAM1 on cytokine-activated ECs. The pH-responsive delivery of VCAM1 siRNA efficiently reduced temporal surface protein expression, which may be used to avert leukocyte recruitment.