Gene therapy has attracted much attention because of its unique mechanism of action, non-toxicity, and good tolerance, which can kill cancer cells without damaging healthy tissues. siRNA-based gene therapy can downregulate, enhance, or correct gene expression by introducing some nucleic acid into patient tissues. Routine treatment of hemophilia requires frequent intravenous injections of missing clotting protein. The high cost of combined therapy causes most patients to lack the best treatment resources. siRNA therapy has the potential of lasting treatment and even curing diseases. Compared with traditional surgery and chemotherapy, siRNA has fewer side effects and less damage to normal cells. The available therapies for degenerative diseases can only alleviate the symptoms of patients, while siRNA therapy drugs can upregulate gene expression, modify epigenetic changes, and stop the disease. In addition, siRNA also plays an important role in cardiovascular diseases, gastrointestinal diseases, and hepatitis B. However, free siRNA is easily degraded by nuclease and has a short half-life in the blood. Research has found that siRNA can be delivered to specific cells through appropriate vector selection and design to improve the therapeutic effect. The application of viral vectors is limited because of their high immunogenicity and low capacity, while non-viral vectors are widely used because of their low immunogenicity, low production cost, and high safety. This paper reviews the common non-viral vectors in recent years and introduces their advantages and disadvantages, as well as the latest application examples.

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Research Status and Prospect of Non-Viral Vectors Based on siRNA: A Review

A temperature-/pH-responsive hydrogel was successfully prepared by in situ free radical polymerization of N-isopropyl acrylamide (NIPAm), itaconic acid (IA), and poly (ethylene glycol) methyl ether acrylate (OEGA). The pore size, mechanical strength, and swelling ratio of the P(NIPAm-IA-OEGA) were strongly affected by the contents of IA. 5-Fluorouracil (5-Fu) was chosen as a model drug, and the maximum drug loading was 68 mg/g. P(NIPAm-IA-OEGA) had a much greater release rate in a simulated intestinal fluid environment (47.73 %, 16 h) than in gastric juice (33.11 %, 16 h). The hemolysis test and cytotoxicity test proved that prepared hydrogel had good biocompatibility. In summary, P(NIPAm-IA-OEGA) can be used for oral drug delivery systems in cancer therapy. 

Synthesis and Characterization of Temperature‐/pH‐Responsive Hydrogels for Drug Delivery

Background Biological ageing is one of the principal risk factors for neurodegenerative diseases. It is becoming increasingly clear that acceleration of DNA methylation age, as measured by the epigenetic clock, is closely associated with many age-related diseases. Methods We searched the PubMed and Web of Science databases to identify eligible studies reporting epigenetic clocks in several neurodegenerative diseases, including Alzheimer’s disease (AD), Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS) and Huntington’s disease (HD). Results Twenty-three studies (12 for AD, 4 for PD, 5 for ALS, and 2 for HD) were included. We systematically summarised the clinical utility of 11 epigenetic clocks (based on blood and brain tissues) in assessing the risk factors, age of onset, diagnosis, progression, prognosis and pathology of AD, PD, ALS and HD. We also critically described our current understandings to these evidences, and further discussed key challenges, potential mechanisms and future perspectives of epigenetic ageing in neurodegenerative diseases. Conclusions Epigenetic clocks hold great potential in neurodegenerative diseases. Further research is encouraged to evaluate the clinical utility and promote the application. PROSPERO registration number CRD42022365233.

Epigenetic clocks in neurodegenerative diseases: a systematic review

Death-associated protein kinase 1 (DAPK1), a Ca2+/calmodulin-regulated serine/threonine kinase, regulates cell apoptosis and autophagy, and has been implicated in the pathogenesis of Alzheimer's disease (AD). Targeting DAPK1 may be a promising approach for treating AD. In our previous study, we found that a natural polyphenol, resveratrol (1), is a moderate DAPK1 inhibitor. In the present study, we investigated the interactions between natural and synthetic derivatives of 1 and DAPK1. Binding assays including intrinsic fluorescence quenching, protein thermal shift and isothermal titration calorimetry indicated that oxyresveratrol (3), a hydroxylated derivative, and pinostilbene (5), a methoxylated derivative, bind to DAPK1 with comparable affinity to 1. The enzymatic assay showed that 3 more effectively inhibits the intrinsic ATPase activity of DAPK1 compared to 1. Crystallographic analysis revealed that the binding mode of the methoxylated derivatives were different from those of 1 and 3, resulting in a unique interaction. Our results suggest that 3 may be helpful in treating AD and provide a clue for the development of promising DAPK1 inhibitors.

Characterization of the Molecular Interactions between Resveratrol Derivatives and Death-associated Protein Kinase 1.

Abstract Acute myeloid leukemia (AML) is a disease of malignant proliferation of abnormally or poorly differentiated myeloid cells of the hematopoietic system. The clinical treatments of non-M3 AML are experiencing a lack of effective drugs. V8 is a newly synthesized derivative of the natural flavonoid wogonin, which belongs to the potential anticancer drug, and has shown significant antitumor activity in vitro and in vivo . In this study, we investigated the effects of V8 on AML cell lines and primary AML cells and the underlying mechanisms. The results showed that V8 inhibited the growth and induced the apoptosis of AML cell lines (ME-1, Kasumi-1, and U937) and primary AML cells in a concentration-dependent manner. Meanwhile, we revealed that V8-induced apoptosis was accompanied by mitochondrial injury, such as the reduction of mitochondrial membrane potential and ATP production, activation of endoplasmic reticulum stress (ERS) characterized by GRP78 and caspase-12 expression upregulation. Mechanism studies have shown that V8 induced mitochondrial injury and inhibited mitophagy via elevating the intracellular ROS level. In addition, V8 activated PERK-p-eIF2α-ATF4 and Ire1α-XBP1 pathways and induced apoptosis of AML cells via selectively activating CHOP. Correspondingly, the degree of apoptosis and expression of apoptosis-related proteins were alleviated after the elimination of cytoplasmic ROS with N-Acetyl-L-cysteine (NAC) and knocking out CHOP in the cells by transfection with CHOP shRNA, implicating that mitochondrial injury-triggered upregulation of ROS and CHOP played an important role in V8-induced apoptosis of AML cells. In primary AML cells-bearing NOD/SCID mice model and U937 cells-inoculating BALB/c nude mice xenografts transplantation tumor model, administration of V8 markedly prolonged survival time and inhibited the xenografts growth via CHOP-mediated ERS in vivo . In conclusion, our study provides a new insight into the mechanism of V8-induced apoptosis, suggesting the potential of V8 as a promising agent against AML. 

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Therapeutic effect of V8 affecting mitophagy and endoplasmic reticulum stress in acute myeloid leukemia mediated by ROS and CHOP signaling

Cellular mitophagy means that cells selectively wrap and degrade damaged mitochondria through an autophagy mechanism, thus maintaining mitochondria and intracellular homeostasis. In recent years, mitophagy has received increasing attention as a research hotspot related to the pathogenesis of clinical diseases, such as neurodegenerative diseases, cardiovascular diseases, cancer, metabolic diseases, and so on. It has been found that the regulation of mitophagy may become a new direction for the treatment of some diseases. In addition, numerous small molecule modulators of mitophagy have also been reported, which provides new opportunities to comprehend the procedure and potential of therapeutic development. Taken together, in this review, we summarize current understanding of the mechanism of mitophagy, discuss the roles of mitophagy and its relationship with diseases, introduce the existing small-molecule pharmacological modulators of mitophagy and further highlight the significance of their development.

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Cellular mitophagy: Mechanism, roles in diseases and small molecule pharmacological regulation

Targeting epigenetics as a promising therapeutic strategy for treatment of neurodegenerative diseases.

Drug carriers can improve the accuracy of drug efficacy and release behaviors. As drug carrier, short peptide hydrogels have promising future since their good biocompatibility and mechanical properties. In this study, we reported a short peptide (FFWDD) that was used as drug carrier to encapsulate doxorubicin (DOX) hydrochloride by forming a drug-loaded hydrogel. The prepared hydrogel has complex interlaced network structure and good mechanical strength that allows it to encapsulate the drug. The strong electrostatic interaction between FFWDD and DOX resulted in less release at pH 7.4 and high release at pH 5.5, which may reduce the side effect of DOX. Drug release kinetics showed release in both pH 7.4 and pH 5.5 is in accordance with the Makoid-Banakar model (R2=0.9973 and 0.9964), reflecting the Non-Fickian diffusion and Quasi-Fickian diffusion release mechanism by the Korsmeyer-Peppas model, respectively. CCK-8 assays demonstrated the excellent biocompatibility of the peptide against human embryonic kidney cells (HEK293) and showed that the antitumor effect of the hydrogel to colon cancer LoVo cells was similar to that of the single DOX. DOX released from the drug delivery system could be taken up by tumor cells in large quantities and accumulate in the nucleus. This research has refined peptide hydrogels as drug carriers, which are expected to be used as drug carriers in biomedical applications. 

Electrostatic Induced Peptide Hydrogels for pH‐Controllable Doxorubicin Release and Antitumor Activity

The death-associated protein kinase (DAPK) family is a member of the calcium/calmodulin-regulated serine/threonine protein kinase family, and studies have shown that its role, as its name suggests, is mainly to regulate cell death. The DAPK family comprises five members, including DAPK1, DAPK2, DAPK3, DRAK1 and DRAK2, which show high homology in the common N-terminal kinase domain but differ in the extra-catalytic domain. Notably, previous research has suggested that the DAPK family plays an essential role in both the development and regulation of human diseases. However, only a few small-molecule inhibitors have been reported. In this Perspective, we mainly discuss the structure, biological function, and role of DAPKs in diseases and the currently discovered small-molecule inhibitors, providing valuable information for the development of the DAPK field.

Targeting Death-Associated Protein Kinases for Treatment of Human Diseases: Recent Advances and Future Directions.

The preparation of polymer therapeutics capable of synergistic co-delivery systems for chemotherapeutic drugs and genes remains a tough problem in cancer therapy. Herein, we develop a novel co-delivery system for chemotherapeutic drugs and genes. Paclitaxel (PTX) was covalently grafted onto the cationic copolymer poly(ethylene glycol)-b-poly(l-lysine) (PEG-b-PLL) via a disulfide bond, and then CXCL1 shRNA was loaded on the cationic micelles PEG-b-PLL-SS-PTX by electrostatic interaction to construct PEG-b-PLL-SS-PTX/pDNA complex. The chemical structure of the complex was confirmed by gel permeation chromatography, 1H NMR and Fourier transform infrared spectroscopy. The binding ability of the cationic micelles and pDNA was detected by gel retardation assay. In vitro gene transfection studies revealed an enhanced gene transfection efficiency due to the co-delivery of PTX and pDNA to ovarian cancer cells. In vitro and in vivo data demonstrated that the co-delivery complex exhibited a synergistic role of chemotherapy and gene therapy and possessed enhanced antitumor effects. The strategy of using drug-loaded cationic micelles as gene delivery carriers is a versatile approach to construct a co-delivery system, which might result in more efficient cancer therapy. © 2022 Society of Industrial Chemistry. 

Yingying Lu

Papers

Cellular mitophagy: Mechanism, roles in diseases and small molecule pharmacological regulation

Targeting epigenetics as a promising therapeutic strategy for treatment of neurodegenerative diseases.

Electrostatic Induced Peptide Hydrogels for pH‐Controllable Doxorubicin Release and Antitumor Activity

Targeting Death-Associated Protein Kinases for Treatment of Human Diseases: Recent Advances and Future Directions.

Co‐delivery of paclitaxel and CXCL1 shRNA via cationic polymeric micelles for synergistic therapy against ovarian cancer