Engineering Bioactive Nanoparticles to Rejuvenate Vascular Progenitor Cells

Fetal exposure to gestational diabetes mellitus (GDM) predisposes children to future health complications including hypertension and cardiovascular disease. A key mechanism by which these complications occur is through stress-induced dysfunction of vascular progenitor cells, including endothelial colony-forming cells (ECFCs). Although several approaches have been previously explored to restore endothelial dysfunction, their widespread adoption remains tampered by systemic side effects of adjuvant drugs and their limited efficacy. Here, we report a strategy to rejuvenate circulating vascular progenitor cells by conjugation of drug-loaded liposomal nanoparticles directly to the surface of GDM-exposed ECFCs (GDM-ECFCs). Bioactive nanoparticles can be robustly conjugated to the surface of ECFCs without altering cell viability and key progenitor phenotypes. Moreover, controlled delivery of therapeutic drugs to vascular progenitor cells is able to normalize transgelin (TAGLN) expression and improve cell migration, which is a critical key step in establishing functional vascular networks. More importantly, sustained pseudo-autocrine stimulation with bioactive nanoparticles is able to improve in vitro and in vivo vasculogenesis of GDM-ECFCs. Collectively, these findings highlight a simple, yet promising strategy to rejuvenate GDM-ECFCs and improve their therapeutic potential, which can be clinically-translated to address various cardiovascular complications, as well as toward a range of approaches in regenerative medicine.

Multifunctional nanoparticle-mediated SHARP1 knockdown in MLL-AF6 acute myeloid leukemia

Acute myeloid leukemia (AML) has an extremely poor prognosis and high relapse and fatality rates. We targeted SHARP1 using multifunctional small interfering RNA (siRNA) and bortezomib (BTZ)-loaded cRGD-guided PEGylated cationic liposomal nanoparticles to monitor their antileukemic activity in MLL-AF6 AML cells. Efficient siRNA/BTZ co-delivery by the nanoparticles significantly inhibited cell viability, decreasing clonogenic growth of AML cells and stimulating robust apoptosis. We hypothesized that SHARP1 downregulation induced nonfunctional MLL-AF6, DOT1L, MEN1, and LEDGF fusion protein accumulation, preventing MLL-AF complex formation and downregulating RAS-GTP and Bcl-2, consequently triggering autophagy and apoptosis. The BTZ combination substantially augmented therapeutic synergy leading to enhanced autophagic and apoptotic events. Our findings demonstrate a state-of-the-art biodegradable nanoplatform for siRNA/BTZ co-delivery with targeted SHARP1 knockdown, demonstrating a potential therapeutic option for MLL-AF6 AML.

The Role of miR-200c Decorated Liposomal Nanoparticles as a Carrier Target for Wnt/β-Catenin Signaling to Regulate Triple-Negative Breast Cancer Cell Line

Treatment of triple-negative breast cancer (TNBC) is not effective and prognosis is poor. This study explored the effect of miR-200c on triple-negative breast cancer cell line (TNBCC) and possible mechanism. L533 cell line was assigned into blank group, empty vector group (transfection of liposome nanoparticles), XAV939 group (transfected with Wnt/β-catenin signaling inhibitor XAV939), miR-200c group (transfection with liposomal nanoparticles as carrier miR-200c), followed by analysis of cell behaviors, Wnt/β-catenin signaling activation and target genes (c-myc, cyclinD1, MMP-2). The cell growth from 1st to 2nd day did not change significantly (P > 0.05). When the cells grew to 3rd day, viable cells in miR-200c group and XAV939 group were significantly lower than in the other two groups. XAV939 and miR-200c groups had highest cell number in G0/G1 phase and lowest cell number in S and G2/M phases along with lowest number of cells passing through matrix membrane (81.47±6.54) and (79.76±6.72) respectively. The expressions of Wnt, β-catenin, c-myc, cyclinD1, MMP-2 protein in XAV939 group and miR-200c group were significantly decreased (P > 0.05). miR-200c can target Wnt/β-catenin signaling. The liposomal nanoparticles can act as carrier for miR and both 200c and XAV939 can inhibit TNBCC behaviors possibly through inhibition of Wnt/β-catenin signaling activity and down-regulation of c-myc, cyclinD1, and MMP-2.

Effects of Liposomal Nanoparticles-Mediated miR-126 on Cervical Cancer Cells via Anti-Programmed Cell Death-1/Programmed Death Ligand 1 (PD-1/PD-L1) Signaling

Cervical cancer is often treated with surgery, radiotherapy and chemotherapy, but it does not have the advantages of precise treatment and prognosis is not ideal. Molecular targeted therapy can make up for the above shortcomings. This study mainly analyzed the influence of miR-126 on cervical cancer cells and possible molecular mechanisms, so as to provide a reference for better clinical improvement of prognosis for cervical cancer. C33a cells were assigned into control group, empty carrier group (C33a cells were co-cultured with liposome nanoparticle carrier), inhibitor group (C33a cells were treated with PD-1/PD-L1 signaling pathway inhibitor), miR-126 group (miR-126 with liposomal nanoparticles as carrier was added to C33a cells), followed by expression analysis of miR-126 and AK2, cell proliferation, PD-1/PD-L1 signaling and phosphorylation levels, as well as tumor mass and volume in nude mice. At 24 h, no difference of cell proliferation was found (P > 0.05) but cell proliferation showed significant differences after cell growth of 48 h, with lower proliferation in inhibitor group and miR-126 group (P < 0.05). The levels of PD-1, PD-L1, AK2, and p-PD-1 in inhibitor group and miR-126 group were significantly lower than for the other two groups (P > 0.05). There was a target relationship between miR-126 and AK2. The transplanted tumor in the miR-126 group was significantly decreased, with lower tumor mass and volume than control group (P < 0.05). The carrier-based miR-126 and PD-1/PD-L1 signaling inhibitors can inhibit cervical cancer cell proliferation.

Exploring molecular mechanism of novel liposomal nanoparticles application in glioma

This study investigated whether miR-509 plays a role in regulating autophagy and apoptosis-related caspase 3 genes, and analyzes targeted nanoparticles intervention in glioma cells. The surgically resected glioma tissue specimens were included as observation group, and control group used a 2-cm open tissue next to the glioma followed by analysis of miR-509 and caspase 3 level by qPCR. Glioma cell line U251 was divided into miRNC group, targeted nanoparticle group, siRNA-NC group, and siRNA-caspase 3 group, followed by analysis of caspase 3 expression, cell proliferation by flow cytometry, and cell invasion and metastasis by trans well. Caspase 3 mRNA expression was significantly higher in glioma tissues compared with controls. Lower miR-509 and higher caspase 3 expression were correlated with TNM stage. Caspase 3 mRNA level was significantly higher and miR-509 was lower in glioma cells or glioma ell line U251 than those in normal cells. Transfection of siRNA-caspase 3, targeted nanoparticles effectively reduced cell proliferation, metastasis, and invasion and down-regulated caspase 3 levels in U251 cells. Reduced miR-509 expression was associated with elevated caspase 3 expression and enhanced invasive metastatic capacity of glioma cells. Overexpression of miR-509 can effectively reduce cell proliferation, metastasis, and invasion by targeted nanoparticles inhibiting caspase 3 expression.