RESUMO
The current research models of breast cancer are usually limited in their capacity to recapitulate the tumor microenvironment in vitro. The lack of an extracellular matrix (ECM) oversimplifies cell-cell or cell-ECM cross-talks. Moreover, the lack of tumor-associated macrophages (TAMs), that can comprise up to 50% of some solid neoplasms, poses a major problem for recognizing various hallmarks of cancer. To address these concerns, a type of direct breast cancer cells (BCCs)-TAMs coculture organoid model was well developed by a sequential culture method in this study. Alginate cryogels were fabricated with appropriate physical and mechanical properties to serve as an alternative ECM. Then, our previous experience was leveraged to polarize TAMs inside of the cryogels for creating an in vitro immune microenvironment. The direct coculture significantly enhanced BCCs organoid growth and cancer aggressive phenotypes, including the stemness, migration, ECM remodeling, and cytokine secretion. Furthermore, transcriptomic analysis and protein-protein interaction networks implied certain pathways (PI3K-Akt pathway, MAPK signaling pathway, etc.) and targets (TNF, PPARG, TLR2, etc.) during breast cancer progression in a TAM-leading immune microenvironment. Future studies to advance treatment strategies for BCC patients may benefit from using this facile model to reveal and target the interactions between cancer signaling and the immune microenvironment.
Assuntos
Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/patologia , Macrófagos Associados a Tumor/metabolismo , Técnicas de Cocultura , Biomimética , Criogéis/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Macrófagos/metabolismo , Microambiente Tumoral , Linhagem Celular TumoralRESUMO
Clinical evidence supports a role for the extracellular matrix (ECM) in cancer plasticity across multiple tumor types. The lack of in vitro models that represent the native ECMs is a significant challenge for cancer research and drug discovery. Therefore, a major motivation for developing new tumor models is to create the artificial ECM in vitro. Engineered biomaterials can closely mimic the architectural and mechanical properties of ECM to investigate their specific effects on cancer progression, offering an alternative to animal models for the testing of cancer cell behaviors. In this review, we focused on the biomaterials from different sources applied in the fabrication of the artificial ECM and their biophysical cues to recapitulate key features of tumor niche. Furthermore, we summarized how the distinct biophysical cues guided cell behaviors of cancer plasticity, including morphology, epithelial-to-mesenchymal transition (EMT), enrichment of cancer stem cells (CSCs), proliferation, migration/invasion and drug resistance. We also discuss the future opportunities in using the artificial ECM for applications of tumorigenesis research and precision medicine, as well as provide useful messages of principles for designing suitable biomaterial scaffolds.
RESUMO
Image 1.
RESUMO
The immunologic response is considered to play a pivotal role in the application of biomaterial implants, and intrinsic properties of biomaterials can significantly modulate the anti-inflammatory effects. However, how physical confinement influences M2 polarization of macrophages and the relevant mechanisms have not been clearly elucidated. In this study, pore size and porosity in cryogels can be mediated by utilizing alginates with different viscosities. Cryogels of small pore size and low porosity can restrict M2 polarization of macrophages in vitro, judging from cell morphology, secretion of cytokines and expression of key M2-related genes. In comparison, cryogels of large pore size or high porosity can induce M2 polarization in vivo, resulting in the anti-inflammation effects. High-throughput RNA-seq analysis demonstrates that the mRNA surveillance pathway is key in the polarization process, and four primary transcription factors (PPAR-γ, STAT6, NF-κB, and STAT1) participate probably by competition in DNA binding to regulate M2-related gene expression. This study confirms that enough physical space inside is necessary to promote M2 polarization for the anti-inflammatory performance, which can be applied widely in the fields of tissue engineering and regenerative medicine.
Assuntos
Alginatos , Criogéis , Anti-Inflamatórios , Criogéis/metabolismo , Macrófagos/metabolismo , Fenótipo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Postsurgical localized chemotherapy for breast cancer recurrence (BCR) still faces many problems which dampen researchers' enthusiasm and discounted prognosis. Simple strategies with controllable toxicities are expected to address these hurdles. Lentinan (LNT) has excellent biocompatibility and notable antitumor activity but rather low bioavailability after intravenous or oral administration. Here, a sponge-like LNT/chitosan composite (LNT/CS sponge) was prepared for efficient local delivery to prevent postoperative BCR. The obtained sponges exhibit uniform porosity and sustained release of LNT in vitro and in vivo. Furthermore, the sponges were implanted and showed significant reduction of postsurgical recurrence and suppression of long-term tumor regrowth with favorable biocompatibility in a subcutaneous postsurgical recurrence mouse model. Subsequent studies revealed that LNT can restrain the stemness of breast cancer cells, which may account for the long-term inhibition of tumor relapse. Therefore, LNT/CS sponge has a great potential as a promising alternative for postsurgical BCR.
Assuntos
Antineoplásicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Quitosana , Lentinano/administração & dosagem , Cuidados Pós-Operatórios , Animais , Antineoplásicos/química , Neoplasias da Mama/terapia , Linhagem Celular Tumoral , Sobrevivência Celular , Quitosana/química , Modelos Animais de Doenças , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Lentinano/química , Camundongos , Recidiva , Análise Espectral , Resultado do Tratamento , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The locoregional recurrence of breast cancer after tumor resection represents several clinical challenges, and conventional post-surgical adjuvant therapeutics always bring about significant systemic side effects. Thus, the local therapy strategy has received considerable interest in breast cancer treatment, and hydrogels can function as ideal platforms due to their remarkable properties such as good biocompatibility, biodegradability, flexibility, and multifunctionality. The nano-hydrogel composites can further incorporate the advantages of nanomaterials into the hydrogel system, to fabricate hierarchical structures for stimulating controlled multi-stage release of different therapeutic agents and improving the synergistic effects of combination therapy. In this review, the problems of clinical treatments of breast cancer and properties of hydrogels in current biomedical applications are briefly overviewed. The focus is on recent advances in local therapy based on nano-hydrogel composites for both monotherapy (chemotherapy, photothermal and photodynamic therapy) and combination therapy (dual chemotherapy, photothermal chemotherapy, photothermal immunotherapy, radio-chemotherapy). Moreover, the challenges and perspectives in the development of advanced nano-hydrogel systems are also discussed.
Assuntos
Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Hidrogéis/química , Nanocompostos/química , Animais , Feminino , Humanos , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
The extracellular matrix (ECM) stiffening is an important sign of local microenvironment change, which is considered as a hallmark of many diseases including hepatocellular carcinoma (HCC). The fates of both cancer cells and immune cells can be regulated by mechanical feedbacks acquired from ECM, but there is a lack of a precise study of mechanical feedback modes in different cell phenotypes following with the progressively increasing ECM stiffness. Herein, we used a biopolymeric film without further modification of adhesive molecules, as a natural local niche to mimic a gradually stiffening manner from HCC onset in liver cirrhosis to its metastasis in the spinal cord. Three distinct manners of mechanical feedbacks were found: the gradual manner in HCC cell spreading, migration and early apoptosis to oxaliplatin, the stepwise manner in HCC cell adhesion, proliferation, focal adhesion (FA) formation, drug resistance, and macrophage M1 polarization; the specific manner in the stages of the progression of epithelial-mesenchymal transition at different stiffness ranges. Further investigation of molecular mechanisms confirmed those mechanical feedback manners by signaling activation of FA kinase, phosphatidylinositol 3-kinase, and expression of pro-/antiapoptotic and pro-/anti-inflammatory genes. Our results pave a novel avenue to know about mechanical feedbacks from ECM, which could be used for future cancer studies and in vitro drug screening applications.
Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Biomimética , Carcinoma Hepatocelular/genética , Retroalimentação , Humanos , Neoplasias Hepáticas/genética , Macrófagos , Fenótipo , Microambiente TumoralRESUMO
Cancer progression is regulated by multiple factors of extracellular matrix (ECM). Understanding how cancer cells integrate multiple signaling pathways to achieve specific behaviors remains a challenge because of the lack of appropriate models to copresent and modulate ECM properties. Here we proposed a strategy to build a thin biomaterial matrix by poly(l-lysine) and hyaluronan as an artificial stiffness-tunable ECM. Transforming growth factor-beta 1 (TGF-ß1) was used as a biochemical cue to present in an immobilized and spatially controlled manner, with a high loading efficiency of 90%. Either soft matrix with immobilized TGF-ß1 (i-TGF) or bare stiff matrix could only promote HCC cells to form the epithelial phenotype, whereas stiff matrix with i-TGF was the only condition to induce the mesenchymal phenotype. Further investigation revealed that i-TGF increased the specific TGF-ß1 receptor (TßRI) expression to activate PI3K pathway. i-TGF-TßRI interactions also promoted HCC cell adhesion to enlarge contact area for stiffness sensing, resulting in the raising expression of the mechano-sensor (ß1 integrin). Mechanotransduction would then be enhanced by the ß1 integrin/vinculin/p-FAK pathway, leading to a noble PI3K activation. Using our model, a novel mechanism was discovered to elucidate regulation of cell fates by coupling mechanotransduction and biochemical signaling.
Assuntos
Carcinoma Hepatocelular , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Matriz Extracelular/química , Neoplasias Hepáticas , Transdução de Sinais/efeitos dos fármacos , Fator de Crescimento Transformador beta1 , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Células Hep G2 , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Proteínas de Neoplasias/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fator de Crescimento Transformador beta1/química , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
Hepatocellular carcinoma develops within an altered mechanical and biochemical environment. The chemokine C-X-C motif chemokine ligand 12 plays an important role in tumor cell metastasis during the process of the epithelial-mesenchymal transition. Here, we successfully engineered a biomimetic matrix made up of polyelectrolyte films with appropriate stiffness, which could present C-X-C motif chemokine ligand 12 in an immobilized status in a spatially controlled manner. The adsorbed amounts of C-X-C motif chemokine ligand 12 could be precisely adjusted over a large range from 27 ng/cm2 to 2.6 µg/cm2. Immobilized C-X-C motif chemokine ligand 12 induced an obvious increase in hepatocellular carcinoma cell adhesion, spreading, and migration in a dose-dependent manner. The phenomenon was associated with proportion changes of three phenotypes of cell morphology (round, polygonal, and elongated) and higher concentrations of immobilized C-X-C motif chemokine ligand 12 led to the rising number of elongated cells along with up-regulation of the mesenchymal markers, as a critical step in epithelial-mesenchymal transition. CD44 and integrin αvß3 revealed the spatial coincidence in three cell phenotypes. Immobilized C-X-C motif chemokine ligand 12-induced ERK phosphorylation also exhibited dose-dependent manner. Our results highlight that an engineered matrix to locally deliver chemokine exhibits the potential to manipulate important cellular responses (including adhesion, spreading, migration, morphology, distribution of surface receptors, and signaling pathways) in the epithelial-mesenchymal transition process during hepatocellular carcinoma development.
Assuntos
Materiais Biomiméticos/química , Carcinoma Hepatocelular/patologia , Quimiocina CXCL12/administração & dosagem , Preparações de Ação Retardada/química , Transição Epitelial-Mesenquimal , Neoplasias Hepáticas/patologia , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Quimiocina CXCL12/farmacologia , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Humanos , Polieletrólitos/químicaRESUMO
The particular characteristics of the tumor microenvironment have the potential to strongly promote tumor growth, metastasis and angiogenesis and induce drug resistance. Therefore, the development of effective, systemic therapeutic approaches specifically based on the tumor microenvironment is highly desirable. Hypoxia-inducible factor-1α (HIF-1α) is an attractive therapeutic target because it is a key transcription factor in tumor development and only accumulates in hypoxic tumors. We report here that a cationic mixed micellar nanoparticle (MNP) consisting of amphiphilic block copolymers poly(ε-caprolactone)-block-poly(2-aminoethylethylene phosphate) (PCL(29)-b-PPEEA(21)) and poly(ε-caprolactone)-block-poly(ethylene glycol) (PCL(40)-b-PEG(45)) was a suitable carrier for HIF-1α siRNA to treat hypoxic tumors, which showed an average diameter of 58.0 ± 3.4 nm. The complex MNP(siRNA), formed by the interaction of MNP and siRNA, was transfected into PC3 prostate cancer cells efficiently, while the inhibition of HIF-1α expression by MNP loaded with HIF-1α siRNA (MNP(siHIF)) blocked PC3 cell proliferation, suppressed cell migration and disturbed angiogenesis under in vitro hypoxic mimicking conditions. It was further demonstrated that systemic delivery of MNP(siHIF) effectively inhibited tumor growth in a PC3 prostate cancer xenograft murine model without activating innate immune responses. Moreover, delivery of MNP(siHIF) sensitized PC3 tumor cells to doxorubicin chemotherapy in vitro and in vivo by downregulating MDR1 gene expression which was induced by hypoxia. The underlying concept of use of MNP(siHIF) to block HIF-1α holds promise as an example of a clinical approach using specific siRNA therapy for cancer treatment aimed at the hypoxic tumor microenvironment.
Assuntos
Hipóxia Celular/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Nanopartículas/administração & dosagem , Neoplasias da Próstata/genética , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , Subfamília B de Transportador de Cassetes de Ligação de ATP , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Inibidores da Angiogênese/administração & dosagem , Inibidores da Angiogênese/química , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Proliferação de Células/efeitos dos fármacos , Doxorrubicina/farmacologia , Inativação Gênica , Humanos , Masculino , Camundongos , Camundongos Nus , Micelas , Nanopartículas/química , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/genética , Tamanho da Partícula , Polímeros/administração & dosagem , Polímeros/química , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , RNA Interferente Pequeno/química , Transfecção/métodos , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/genéticaRESUMO
The hyperbranched poly(amido amine) nanoparticles (HPAMAM NPs) with multiple functions, such as biodegradability, autofluorescence, and specific affinity, were successfully prepared by Michael addition dispersion polymerization of CBA, AEPZ, and N-galactosamine hydrochloride (or N-glucosamine hydrochloride) in a mixture of methanol/water. The resultant NPs displayed strong photoluminescence, high photostability, broad absorption, and emission (from 430 to 620 nm) spectra. The fluorescence from HPAMAM NPs may be attributed to the tertiary amine chromophore. The incubation results of the liver cancer cells, HepG2, with the NPs showed that the NPs are nontoxic and can be recognized by asialoglycoprotein receptors on the surface of HepG2 and then can be internalized. Therefore, they have potential applications in bioimaging and drug or gene delivery.
Assuntos
Nanopartículas , Poliaminas/síntese química , Espectroscopia de Ressonância Magnética , Poliaminas/química , Espectrometria de FluorescênciaRESUMO
MiRNAs are viable therapeutic targets for cancer therapy, but the targeted delivery of miRNA or its anti-miRNA antisense oligonucleotides (AMOs) remains a challenge. We report here a PEGylated LPH (liposome-polycation-hyaluronic acid) nanoparticle formulation modified with cyclic RGD peptide (cRGD) for specific and efficient delivery of AMO into endothelial cells, targeting α(v)ß3 integrin present on the tumor neovasculature. The nanoparticles effectively delivered anti-miR-296 AMO to the cytoplasm and downregulated the target miRNA in human umbilical vein endothelial cells (HUVECs), which further efficiently suppressed blood tube formulation and endothelial cell migration, owing to significant upregulation of hepatocyte growth factor-regulated tyrosine kinase substrate (HGS), whereas nanoparticles without cRGD modification showed only little AMO uptake and miRNA silencing activity. In vivo assessment of angiogenesis using Matrigel plug assay also demonstrated that cRGD modified LPH nanoparticles have potential for antiangiogenesis in miRNA therapeutics. With the delivery of anti-miR-296 AMO by targeted nanoparticles, significant decrease in microvessel formulation within Matrigel was achieved through suppressing the invasion of CD31-positive cells into Matrigel and prompting HGS expression in angiogenic endothelial cells.
Assuntos
MicroRNAs/genética , Nanopartículas/química , Oligonucleotídeos Antissenso/genética , Oligopeptídeos/química , Animais , Western Blotting , Linhagem Celular , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Feminino , Humanos , Lipossomos/química , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Modelos Teóricos , Fosfoproteínas/genética , Fosfoproteínas/metabolismoRESUMO
A brush-shaped polymer PHEMA-g-(PEI-b-PEG) with poly(2-hydroxyethyl methacrylate) (PHEMA) backbone and linear poly(ethylenimine)-b-poly(ethylene glycol) (PEI-b-PEG) side chains was synthesized and evaluated as a vector for potential cancer gene therapy. The characterizations by (1)H NMR and laser light scattering demonstrated the brush structure of the polymer. PHEMA-g-(PEI-b-PEG) was much less cytotoxic when compared with branched poly(ethylenimine) with M(w) of 25 kDa. The capacity of plasmid DNA condensation by PHEMA-g-(PEI-b-PEG) was demonstrated by gel retardation assay, and they formed nanosized complexes with surface zeta potential around 20 mV at N/P ratios higher than 5:1. The complexes of PHEMA-g-(PEI-b-PEG) with plasmid DNA were more efficiently internalized by BT474 cells in comparison with the complexes of PEI25K, leading to higher gene transfection in cells. Further investigation using complexes of PHEMA-g-(PEI-b-PEG) with plasmid DNA encoding wild-type p53 gene showed its potential as a carrier for cancer gene therapy. The complexes of PHEMA-g-(PEI-b-PEG) successfully induced elevated wild-type p53 expression in BT474 cells and led to enhanced apoptosis of BT474 cells. Transfection of wild-type p53 using the complexes also significantly increased the sensitivity of BT474 cells to doxorubicin chemotherapy, suggesting the potential of this carrier in cancer gene therapy.
Assuntos
DNA/administração & dosagem , Técnicas de Transferência de Genes , Genes p53 , Poliaminas/química , Polietilenoglicóis/química , Polietilenoimina/análogos & derivados , Poli-Hidroxietil Metacrilato/análogos & derivados , Poli-Hidroxietil Metacrilato/química , Animais , Antibióticos Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA/genética , Doxorrubicina/farmacologia , Humanos , Camundongos , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Tamanho da Partícula , Polieletrólitos , Polietilenoglicóis/efeitos adversos , Polietilenoglicóis/síntese química , Polietilenoimina/química , Poli-Hidroxietil Metacrilato/efeitos adversos , Poli-Hidroxietil Metacrilato/síntese química , Ratos , Ratos Sprague-Dawley , TransfecçãoRESUMO
Aqueous dispersions of thermosensitive gold nanoparticles protected by diblock copolymers of poly(ethylene glycol) and polyphosphoester were prepared and studied. Diblock copolymers MPEG-b-P(EEP-co-PEP) with different compositions that are composed of monomethoxy poly(ethylene glycol), random copolymer of ethyl ethylene phosphate (EEP), and isopropyl ethylene phosphate (PEP) were synthesized by ring-opening polymerization in bulk. Thioctic acid was then conjugated to the terminal hydroxyl group of the polyphosphoester block by esterification. Gold nanoparticles were then prepared by a one-step method and showed core-shell structure with an average gold core diameter of about 10 nm surrounded by a MPEG-b-P(EEP-co-PEP) shell with a thickness of about 30 nm. These polymer stabilized gold nanoparticles are reversibly thermosensitive in aqueous medium, exhibiting tunable collapse temperatures which are dependent on the composition of the diblock copolymers. Methyl tetrazolium (MTT) assay against HEK 293 cells demonstrated that these gold nanoparticles are with good biocompatibility. These gold nanoparticles protected by thermosensitive diblock copolymers with tunable collapse temperature are expected to be useful for biomedical applications.
Assuntos
Ésteres/química , Ouro/química , Nanopartículas/química , Polietilenoglicóis/química , Materiais Biocompatíveis/química , Linhagem Celular , Corantes/farmacologia , Humanos , Espectroscopia de Ressonância Magnética/métodos , Modelos Químicos , Nanoestruturas/química , Nanotecnologia/métodos , Polímeros/química , Temperatura , Sais de Tetrazólio/farmacologia , Tiazóis/farmacologiaRESUMO
Cellular specific micellar systems from functional amphiphilic block copolymers are attractive for targeted intracellular drug delivery. In this study, we developed reactive micelles based on diblock copolymer of poly(ethyl ethylene phosphate) and poly(epsilon-caprolactone). The micelles were further surface conjugated with galactosamine to target asialoglycoprotein receptor (ASGP-R) of HepG2 cells. The size of micellar nanoparticles was about 70nm in diameter, and nanoparticles were negatively charged in aqueous solution. Through recognition between galactose ligands with ASGP-R of HepG2 cells, cell surface binding and internalization of galactosamine-conjugated micelles were significantly promoted, which were demonstrated by flow cytometric analyses using rhodamine 123 fluorescent dye. Paclitaxel-loaded micelles with galactose ligands exhibited comparable activity to free paclitaxel in inhibiting HepG2 cell proliferation, in contrast to the poor inhibition activity of micelles without galactose ligands particularly at lower paclitaxel doses. In addition, population of HepG2 cells arrested in G2/M phase was in positive response to paclitaxel dose when cells were incubated with paclitaxel-loaded micelles with galactosamine conjugation, which was against the performance of micelles without galactose ligand, owing to the ligand-receptor interaction. The surface functionalized micellar system is promising for specific anticancer drug transportation and intracellular drug release.