Establishment of a hTfR mAb-functionalized HPPS theranostic nanoplatform.

Rational: Many efforts have been made to develop ligand-directed nanotheranostics in cancer management which could afford both therapeutic and diagnostic functions as well as tumor-tailored targeting. Theranostic nanoplatform targeting transferrin receptor (TfR) is an effective system for favorable delivery of diagnostic and therapeutic agents to malignancy site. Methods: To enable amalgamation of therapy and diagnosis to many TfR+ tumor, hTfR (human TfR) monoclonal antibody (mAb)-functionalized HPPS nanoparticle (HPPS-mAb) was prepared with hTfR mAb on the shell and with fluorophore DiR-BOA in the core. The targeting specificity was investigated in vitro by immunostaining and in vivo using a double-tumor-engrafted mouse model. HPPS-mAb/siRNA effect on HepG2 cells was determined by RT-PCR and western blot. Results: HPPS-mAb could specifically target cancer cells through TfR and achieve tumor accumulation at an early valuable time node, thus efficiently delivering therapeutic survivin siRNA into TfR+ HepG2 cells and mediating cell apoptosis. DiR-BOA can act as an imaging tool to diagnose cancer. Conclusions: Our studies provide a promising TfR mAb-directed theranostic nanoplatform candidate in tumor molecular imaging and in TfR targeted tumor therapy.

Selection for Anti-transferrin Receptor Bispecific T-cell Engager in Different Molecular Formats.

Selecting an ideal molecular format from diverse structures is a major challenge in developing a bispecific antibody (BsAb). To choose an ideal format of anti-CD3 × anti-transferrin receptor (TfR) bispecific antibodies for clinical application, we constructed TfR bispecific T-cell engager (BiTE) in two extensively applied formats, including single-chain tandem single-chain variable fragments (scFvs) and double-chain diabodies, and evaluated their functional characterizations in vitro. Results demonstrated that TfR-BiTE in both formats directed potent killing of TfR+ HepG2 cells. However, compared to two-chain diabodies, scFvs were more efficient in antigen binding and TfR+ target killing. Furthermore, different domain orders in scFvs would also be evaluated because single-TfR-CD3-His was preferable to single-CD3-TfR-His in immunotherapeutic strategies. Thus, the single-chain tandem TfR-CD3 format was favored for further investigation in cancer therapy.

Therapeutic Bispecific T-Cell Engager Antibody Targeting the Transferrin Receptor.

Bispecific T-cell engager antibodies (BiTE) have been explored as a means to recruit cytolytic T cells to kill tumor cells. The transferrin receptor (TfR) is highly expressed on the surface of rapidly proliferating tumor cells. Therefore, it holds great potential in T cell redirecting therapies. In this research, we developed a BiTE targeting TfR and CD3 (TfR-BiTE) and studied its therapeutic impact on TfR-positive cancer. TfR-BiTE had the ability to induce the selective lysis of various TfR-positive cancer cells through the activation of T cells, the release of cytokines, and then the coming proliferation of T cells, whereas TfR-negative cells were not affected. In a subcutaneous HepG2 xenograft model, low concentrations of TfR-BiTE inhibited tumor growth. Overall, these results reveal that TfR-BiTE can selectively deplete TfR-positive HepG2 cells; hence, it represents a novel immunotherapeutic approach for the treatment of hepatocellular carcinoma.

Multiwalled Carbon Nanotubes Prevent Tumor Metastasis Through Switching M2-Polarized Macrophages to M1 via TLR4 Activation.

Targeting tumor-associated macrophages (TAMs) has emerged as a novel therapeutic strategy for cancer metastasis. Here, we investigated whether carboxylated multiwalled carbon nanotubes (MWCNTs-COOH) prevent tumor metastasis through regulating macrophage polarization. In Lewis lung carcinoma (LLC) or B16F10 melanoma-bearing mice, intratracheal instillation of MWCNTs-COOH significantly reduced metastatic burden in the lungs. MWCNTs-COOH promoted the expression of M1 markers (iNOS and CXCR10) and inhibited the expression of M2 markers (CD206 and Arg-1) along with an increased expression of Th1 cytokines (TNF-α and IL-12) and decreased expression of Th2 cytokines (TGF-ß and IL-10). Such changes were accompanied with TLR4 mRNA and protein elevation. Importantly, macrophage depletion in mice lungs reversed the anti-metastatic effects of MWCNTs-COOH. In vitro, MWCNTs-COOH switched IL-4/13-treated macrophages to the M1 phenotype and thus prevented the migration and invasion of LLC cells, accompanied by the upregulation of toll-like receptor (TLR)-4/NF-κB p65 signaling. Moreover, TAK-242 (resatorvid), a specific TLR4 inhibitor, reversed the effects of MWCNTs-COOH on macrophage polarization. In summary, MWCNTs-COOH effectively prevent tumor metastasis through skewing M2-polarized macrophages to M1 via activating TLR4/NF-κB signaling. Thus, targeting TAMs by MWCNTs-COOH is a potential therapeutic approach against tumor metastasis.

Selection of key sequence-based features for prediction of essential genes in 31 diverse bacterial species.

Genes that are indispensable for survival are essential genes. Many features have been proposed for computational prediction of essential genes. In this paper, the least absolute shrinkage and selection operator method was used to screen key sequence-based features related to gene essentiality. To assess the effects, the selected features were used to predict the essential genes from 31 bacterial species based on a support vector machine classifier. For all 31 bacterial objects (21 Gram-negative objects and ten Gram-positive objects), the features in the three datasets were reduced from 57, 59, and 58, to 40, 37, and 38, respectively, without loss of prediction accuracy. Results showed that some features were redundant for gene essentiality, so could be eliminated from future analyses. The selected features contained more complex (or key) biological information for gene essentiality, and could be of use in related research projects, such as gene prediction, synthetic biology, and drug design.

[The effects of si-wu-tang on serum protein of blood deficient mice induced by radiation].

OBJECTIVE: To study the effects of Si-Wu-Tang on serum protein of blood deficient mice b y proteomicstechnique and study the enriching and regulating blood mechanism of Si-Wu-Tang on mocular level. METHOD: The blood deficient mice was induced by using a single dose of 3.5 Gy radiation from a 60Cogamma source, and high resolution two-dimensional polyacryamide gel electrophoresis (2-DE), computer-assisted image analysis, and mass spectrometry were used to detect regulated protein by Si-Wu-Tang. RESULT: 12 lower and 4 higher protein in sera could be recovered by Si-Wu-Tang, 4 protein might be DNA-dependent protein kinase catalytic subunit, Dystrophin, KIF13A, dystonin. They play a part in DNA double-stranded break repair, recombination and modulation of transcription, transportation of mannose-6-phosphate receptor, etc. CONCLUSION: Si-Wu-Tang can regulate serum protein in blood deficient mice, resulting in improving hematopoiesis and lessening irradiated injury.