Gene Therapy Strategies Research Articles

Dual‐functional Nanosized Albumin Facilitates CRISPR‐Cas9‐PAR2 to Mitigate Cytokine Storm via TLR4 and Pyroptosis Signaling

AbstractProtease‐activated receptor 2 (PAR2) widely modulates various cytokine secretions and inflammatory responses, while excessive cytokine releases and immune hyperactivation may trigger cytokine storm. However, the potential participation of PAR2 in cytokine storm remains elusive. CRISPR‐Cas9 as an efficient gene editing tool can be used for investigating the possible function of PAR2. In this study, albumin‐based nanoparticles (hAlbumin/cgPAR2) to protect and deliver CRISPR‐Cas9 for precise gene editing of PAR2 is constructed. Nanosized albumin largely assisted CRISPR‐Cas9‐PAR2 (cgPAR2) to enter cells via scavenger receptor‐mediated endocytosis, and then triggered potent transfection efficiency, consequently stimulating PAR2 knockout in vitro and in vivo. Apart from delivering CRISPR‐Cas9, nanosized albumin could be colocalized with TLR4 on endosome and promote PAR2 deficiency to attenuate TLR4/NF‐κB signaling, exerting the strong inhibitory effect of albumin in inflammatory responses depending on TLR4. Moreover, PAR2 deficiency induced by hAlbumin/cgPAR2 alleviated ERK/STING/NLRP3 signaling and GSDMD‐mediated pyroptosis, consequently inhibiting cytokine storm. Therefore, this study uncovered dual functions of nanosized albumin in delivering CRISPR‐Cas9 and facilitating PAR2 deficiency to prevent cytokine storm. It also uncovered that the significant role of PAR2 in modulating cytokine storm and provided a potential gene therapy strategy for treating this disease.

Role of RET and BRAF genes in thyroid cancer with relevance in gene therapy treatment

At present, cancer is a global problem, among all the types that exist, one of those that was taken into account for this research is thyroid cancer, especially associated with the predisposing genes of RET rearrangement and BRAF gene mutation, which incite the rapid exit of papillary thyroid carcinoma, from MAPK signaling pathways and conformational change by mutation in BRAF at position (V600E) and (V599E). The aim of the subsequent study is to provide recent information on the genes involved, as well as the potential impact of targeted therapies to inhibit these pathways, such as selective RET and BRAF inhibitors, providing a strategy for gene therapy and rather than the traditional approach of thyroidectomy. In addition to the way of its diagnosis by means of different genetic markers, oncogenic genes, genome sequence and generalized findings of the endocrine-genetic problem. This research also explores non-specific associations of the above-mentioned causes, generating a broader knowledge for future medical and genetic adaptation of the disease.

Myopia in Children: Epidemiology, Genetics, and Emerging Therapies for Treatment and Prevention

Refractive errors, particularly myopia, are among the most prevalent visual impairments globally, with rising incidence in children and adolescents. This review explores the epidemiology and risk factors associated with the development of refractive errors, focusing on the environmental and lifestyle factors contributing to the current surge in myopia. We provide an overview of key genetic factors and molecular pathways driving the pathogenesis of myopia and other refractive errors, emphasizing the complex interplay between genetic predisposition and environmental triggers. Understanding the underlying mechanisms is crucial for identifying new strategies for intervention. We discuss current approaches to slow myopia progression in pediatric populations, including pharmacological treatment regimens (low-dose atropine), optical interventions, and lifestyle modifications. In addition to established therapies, we highlight emerging innovations, including new pharmacological agents and advanced optical devices, and insights into potential future treatments. Cutting-edge research into gene therapy, molecular inhibitors, and neuroprotective strategies may yield novel therapeutic targets that address the root causes of refractive errors. This comprehensive review underscores the importance of early intervention and highlights promising avenues for future research, aiming to provide pediatricians with guidance to ultimately improve clinical outcomes in managing and preventing myopia progression in children and young adults.

SNA·SMNP·CBE system: A novel integrative strategy for β-hemoglobinopathies gene therapy

Here, we developed and demonstrated a novel integrative system—Silica Nanorods (SNA) substrate cell capture combined with Supramolecular Nanoparticle (SMNP) delivery mediated CBE base editing (SNA·SMNP·CBE)—achieving the synchronization of CD34+HSPCs cell capture and gene editing for β-hemoglobinopathies. First, in vitro study shows it enables efficient and precise modification of BCL11A promoter in CD34+HSPCs, yielding the highly editing efficiency of 50.4 %, thus making an alternative strategy to conventional immunomagnetic cell separation and electroporation transfection system mediated CBE editing (IMS·EP·CBE). Then, we transplanted the edited human CD34+HSPCs into severe combined immunodeficiency (SCID) mice by using intraosseous injection strategy. When compared with conventional IMS·EP·CBE methods, our results showed that significantly higher human HBG expression in the bone marrow and peripheral blood of recipient mice, and long-term engraftment, evidenced from similar gene expression profiles to naïve CD34+HSPCs at 14 weeks. Conclusively, our integrative system—SNA·SMNP·CBE·intraosseous injection—offers an appealing novel way for the unique potential of gene therapy in the clinic application for β-hemoglobinopathies patients.

Tropism of adeno-associated virus serotypes in mouse lungs via intratracheal instillation

BackgroundGene therapy holds great potential for treating various acquired and inherited pulmonary diseases. Adeno-associated viral (AAV) vectors have been thought to be primary candidates for gene delivery in patients with pulmonary diseases. However, the tropism of AAVs in the lungs remains largely unknown.ResultsHere, we investigate the tropism of twenty serotypes of AAVs by examining AAV-packed vector expression of the enhanced green fluorescent protein (eGFP) in mice. AAV1, AAV4, AAV5, AAV6, AAV6.2, AAV-PHP.B, and AAV-PHP.S exhibit high transduction rates in the airway epithelium. AAV1, AAV4, AAV5, AAV6, and AAV6.2 highly infect club cells. AAV1, AAV4, AAV5, AAV6, AAV6.2, and AAV-PHP.B efficiently infect ciliated cells. AAV8 and AAVrh10 can infect a few alveolar type I cells. AAV1, AAV5, AAV6, AAV6.2, AAV9, and AAVie can infect alveolar type II cells. AAV1, AAV5, AAVie, AAV-PHP.B, AAV-PHP.eB, and AAV-PHP.S can infect a few endothelial cells. However, none of these AAVs can efficiently infect neuroendocrine or smooth muscle cells.ConclusionsOur findings provide comprehensive information about the tropism of AAVs in pulmonary epithelium in mice, which might be helpful in developing efficient AAV-mediated gene therapy strategies for pulmonary disease treatment.

Analysis of the heterologous expression, localization, and cellular response to the Zika virus E protein in vitro.

Zika virus (ZIKV) infection has been associated with damage to neural stem cells in microcephaly in newborns. The virus possesses specific tropism for glioma stem cells mediated by the ZIKV E protein. This infection causes endoplasmic reticulum stress and activation of the unfolded protein response (UPR). However, the cellular response to the expression of the ZIKV E protein alone is unknown. Therefore, in this study, we determined the effect of the expression of the ZIKV E protein on cellular responses and its subcellular localization in HEK-293T cells, due to their use as a biotechnological tool for cellular and lentiviral therapy. We observed that the ZIKV E protein is synthesized in the cytoplasm and inserted into the endoplasmic reticulum (ER), without causing activation of the UPR or cell death, and it is finally transported and located in the cell membrane. Moreover, the expression of the ZIKV E protein does not induce UPR or apoptosis in glioma cells. These results help us to better understand the characteristics of this protein and its possible use as a biotechnological tool for the creation of different gene therapy strategies, vaccines, and synthetic vectors with tropism for neural and glioma stem cells.

Prevalence of Molecular Diagnoses for Usher Syndrome and the Need for Coordinated Care.

Usher syndrome (USH) is a rare, autosomal-recessive genetic disorder and a leading cause of early onset deaf-blindness. A clinical diagnosis is made by the presence of retinitis pigmentosa (RP) with sensorineural hearing loss (SNHL). Subtype (USH1, USH2, USH3) is determined by severity and age of onset. Molecular testing is able to further distinguish USH subtypes by causative gene. As gene therapy strategies continue to be explored for USH, it is important to know the underlying genetic cause and to coordinate care among an interdisciplinary team. We reviewed charts of 198 individuals presenting to the RP clinic at Bascom Palmer Eye Institute (BPEI) for suspected USH. Demographic information, USH clinical diagnosis, molecular testing, molecular diagnosis, and audiological data were collected. Of the 198 patients reviewed, 190 (96%) met clinical criteria for USH and received a clinical diagnosis. There were 67 (35%) that had a genetic test with a pathogenic molecular diagnosis. The average ages at molecular diagnosis were USH1B, 20 years old; USH2A, 37 years old; USH2C, 50 years old. Of the 67 with a molecular diagnosis, 23 (34%) established ophthalmic care and 8 of these (11%) established audiological care. Molecular testing and diagnosis should be part of the routine care of USH individuals to facilitate earlier interventions and coordinated care between ophthalmology and audiology. 4 Laryngoscope, 2024.

EXTH-48. ADENO-ASSOCIATED VIRUS TARGETING OF THE TUMOR MICROENVIRONMENT TO IMPROVE LYMPHOCYTE RECRUITMENT IN GLIOBLASTOMA

Abstract BACKGROUND Glioblastoma (GBM) is notoriously devoid of lymphocytes driven in part by a paucity of lymphocyte trafficking factors necessary to prompt their recruitment, infiltration, and activation. We have developed a novel recombinant adeno-associated virus (AAV) gene therapy strategy that enables focal and stable reconstitution of the GBM tumor microenvironment (TME) with C-X-C motif ligand 9 (CXCL9), a powerful call-and-receive chemokine for cytotoxic T lymphocytes (CTLs). By increasing lymphocyte recruitment in GBM, we HYPOTHESIZE that these tumors will be more responsive to immunotherapy. METHODS 3D fluorescence microscopy was used to characterize the distribution, tropism, and durability of AAV6 transgene expression in preclinical GBM. Flow cytometry and 3D fluorescence microscopy was used to quantify T cell infiltration and visualize their biodistribution in preclinical GBM treated with AAV-CXCL9 in combination with anti-PD-1 immune checkpoint blockade (ICB). Single-cell RNA sequencing was used to characterize intratumor lymphocyte subsets following treatment. Survival studies with and without lymphocyte depletion were done to evaluate immunogenic responsiveness of therapy. RESULTS Focal delivery of CXCL9 with AAV6 results in stable transduction of peri-tumoral astrocytes. When combined with anti-PD-1 ICB, lymphocyte infiltration was improved, with cells found disseminated across the tumor stroma and within the tumor parenchyma. CXCR3, the chemokine receptor for CXCL9, was expressed across lymphocyte subsets and NKT cells, validating treatment selectivity of AAV6-CXCL9 for lymphocytes. Combination treatment considerably improved overall survival in two distinct GBM models, with durable responses dependent on CD8 T-lymphocyte infiltration. CONCLUSIONS By manipulating local chemokine directional guidance, AAV-CXCL9 increases tumor infiltration by CD8-postive cytotoxic lymphocytes, sensitizing GBM to anti-PD-1 ICB. These effects are accompanied by immunologic signatures evocative of an inflamed and responsive TME. These findings support targeted AAV gene therapy as a promising adjuvant strategy for reconditioning GBM immunogenicity given its excellent safety profile, TME-tropism, modularity, and off-the-shelf capability.

Emerging Role of Gene Therapy in Cancer

Cancer is one of the leading causes of death in the world, and researchers have been persistently studying the treatment of cancer. Although traditional treatment methods such as surgery, chemotherapy and radiotherapy are effective, they are often accompanied by serious side effects, drug resistance or limited effects on certain types of cancer. Therefore, it is urgent to develop more accurate and effective treatment plans. Cancer gene therapy is an innovative method to treat or prevent cancer by introducing or editing genetic material. Gene therapy brings new hope to the treatment of cancer, repairing mutant genes, activating the immune system or directly inducing cancer cell death. This review introduces some hot strategies of cancer gene therapy, including suicide gene therapy, tumor suppressor gene activation, immunotherapy, inhibition of oncogene activation, antiangiogenic gene therapy. The principles, application examples and advantages and disadvantages of these therapies in clinical research are also discussed. In addition, the challenges faced by gene therapy in improving safety and reducing side effects, and points out the direction of future research are also described, aiming to promote the transformation of these innovative therapies into conventional clinical applications.

Nanotechnology and CRISPR/Cas-Mediated Gene Therapy Strategies: Potential Role for Treating Genetic Disorders.

Gene therapy has made substantial progress in the treatment of the genetic diseases, focussing on the reduction of characteristics of recessive/dominant disorders, as well as various cancers. Extensive research has been conducted in the past few decades to investigate the application of nanotechnology and CRISPR/Cas technology in gene therapy. Nanotechnology due to attributes such has targeted drug delivery, controlled release, scalability and low toxicity has gained attention of the medical world. CRISPR/Cas9 system is considered as an impactful genome editing tool in the area of next-generation therapeutics and molecular diagnostics. CRISPR technology emphasises on gene editing, gene regulation modulation, and formulation of defined genetic changes. Its applications in treatment of the genetic disorders are extended beyond traditional therapies. These techniques are being explored as treatment of several genetic disorders including Duchenne muscular dystrophy, cystic fibrosis, Alzheimer's disease, Parkinson's disease, and Huntington disease. Despite considerable therapeutic potential of gene therapy, several obstacles must be addressed before it can be widely adopted in clinical practice, particularly in terms of ensuring safety and effectiveness. As research advances in this captivating field, these therapies will become the primary treatments and will have significant beneficial effects on the lives of patients with genetic disorders.

Gene therapy for glaucoma: Targeting key mechanisms

Glaucoma is a group of optic neuropathies characterised by progressive retinal ganglion cell (RGC) degeneration and is the leading cause of irreversible blindness worldwide. Current treatments for glaucoma focus on reducing intraocular pressure (IOP) with topical medications. However, many patients do not achieve sufficient IOP reductions with such treatments. Patient compliance to dosing schedules also poses a significant challenge, further limiting their effectiveness. While surgical options exist for resistant cases, these are invasive and carry risks of complications. Thus, there is a critical need for better strategies to prevent irreversible vision loss in glaucoma. Gene therapy holds significant promise in this regard, offering potential long-term solutions by targeting the disease’s underlying causes at a molecular level. Gene therapy strategies for glaucoma primarily target the two key hallmarks of the disease: elevated IOP and RGC death. This review explores key mechanisms underlying these hallmarks and discusses the current state of gene therapies targeting them. In terms of IOP reduction, this review covers strategies aimed at enhancing extracellular matrix turnover in the conventional outflow pathway, targeting fibrosis, regulating aqueous humor production, and targeting myocilin for gene-specific therapy. Neuroprotective strategies explored include targeting neurotrophic factors and their receptors, reducing oxidative stress and mitochondrial dysfunction, and preventing Wallerian degeneration. This review also briefly highlights key research priorities for advancing gene therapies for glaucoma through the clinical pipeline, such as refining delivery vectors and improving transgene regulation. Addressing these priorities will be essential for translating advancements from preclinical models into effective clinical therapies for glaucoma.

Self‐Initiated Nano‐Micelles Mediated Covalent Modification of mRNA for Labeling and Treatment of Tumors

AbstractAs a promising gene therapy strategy, controllable small molecule‐mRNA covalent modification in tumor cells could be initiated by singlet oxygen (1O2) to complete the modification process. However, in vivo generation of 1O2 is usually dependent on excitation of external light, and the limited light penetration of tissues greatly interferes the development of deep tumor photo therapy. Here, we constructed a tumor‐targeting nano‐micelle for the spontaneous intracellular generation of 1O2 without the need for external light, and inducing a high level of covalent modification of mRNA in tumor cells. Luminol and Ce6 were chemically bonded to produce 1O2 by chemiluminescence resonance energy transfer (CRET) triggered by high levels of hydrogen peroxide (H2O2) in the tumor microenvironment (TME). The sufficient 1O2 oxidized the loaded furan to highly reactive dicarbonyl moiety, which underwent cycloaddition reaction with adenine (A), cytosine (C) or guanine (G) on the mRNA for interfering with the tumor cell protein expression, thereby inhibiting tumor progression. In vitro and in vivo experiments demonstrated that this self‐initiated gene therapy nano‐micelle could induce covalent modification of mRNA by 1O2 without external light, and the process could be monitored in real time by fluorescence imaging, which provided an effective strategy for RNA‐based tumor gene therapy.

Homeostatic regulation of brain activity: from endogenous mechanisms to homeostatic nanomachines.

After the initial concepts of the constancy of the internal milieu or homeostasis, put forward by Claude Bernard and Walter Cannon, homeostasis emerged as a mechanism to control oscillations of biologically meaningful variables within narrow physiological ranges. This is a primary need in the central nervous system that is continuously subjected to a multitude of stimuli from the internal and external environments that affect its function and structure, allowing to adapt the individual to the ever-changing daily conditions. Preserving physiological levels of activity despite disturbances that could either depress neural computation or excessively stimulate neural activity is fundamental, and failure of these homeostatic mechanisms can lead to brain diseases. In this review, we cover the role and main mechanisms of homeostatic plasticity involving the regulation of excitability and synaptic strength from the single neuron to the network level. We analyze the relationships between homeostatic and Hebbian plasticity and the conditions under which the preservation of the excitatory/inhibitory balance fails, triggering epileptogenesis and eventually epilepsy. Several therapeutic strategies to cure epilepsy have been designed to strengthen homeostasis when endogenous homeostatic plasticity mechanisms have become insufficient or ineffective to contrast hyperactivity. We describe "on demand" gene therapy strategies, including optogenetics, chemogenetics, and chemo-optogenetics, and particularly focus on new closed loop sensor-actuator strategies mimicking homeostatic plasticity that can be endogenously expressed to strengthen the homeostatic defenses against brain diseases.

Brain targeted biomimetic siRNA nanoparticles for drug resistance glioblastoma treatment

Glioblastoma multiforme (GBM), the most aggressive intracranial neoplasm, remains incurable at present, primarily due to drug resistance, which significantly contributes to elevated recurrence rates and dismal prognosis. Signal transducer and activator of transcription 3 (STAT3) is a critical gene closely associated with GBM drug resistance and the progression of GBM stem cells (GSCs), making it a promising therapeutic target. In this study, we developed cancer cell membrane-cloaked biomimetic nanoparticles to deliver STAT3 siRNA to reverse drug resistance in homologous GBM. These biomimetic nanoparticles leverage homotypic targeting, rapid endosome escape, and fast siRNA release, leading to efficient in vitro STAT3 knockdown in both temozolomide-resistant U251-TR cells and X01 GSCs. Moreover, benefited from the membrane functionalization, significant prolonged blood circulation, improved blood brain barrier (BBB) penetration and GBM tumor accumulation are achieved by these siRNA biomimetic nanoparticles. Importantly, these nanoparticles effectively inhibit tumor proliferation, significantly extending median survival time in orthotopic U251-TR (43.5 d versus 20 d for PBS control) and X01 GSC-bearing mouse xenografts (52 d versus 19.5 d for PBS control). Altogether, this biomimetic siRNA platform offers a promising strategy for gene therapy targeting drug-resistant GBM.

The Role of Pericytes in Inner Ear Disorders: A Comprehensive Review.

Inner ear disorders, including sensorineural hearing loss, Meniere's disease, and vestibular neuritis, are prevalent conditions that significantly impact the quality of life. Despite their high incidence, the underlying pathophysiology of these disorders remains elusive, and current treatment options are often inadequate. Emerging evidence suggests that pericytes, a type of vascular mural cell specialized to maintain the integrity and function of the microvasculature, may play a crucial role in the development and progression of inner ear disorders. The pericytes are present in the microvasculature of both the cochlea and the vestibular system, where they regulate blood flow, maintain the blood-labyrinth barrier, facilitate angiogenesis, and provide trophic support to neurons. Understanding their role in inner ear disorders may provide valuable insights into the pathophysiology of these conditions and lead to the development of novel diagnostic and therapeutic strategies, improving the standard of living. This comprehensive review aims to provide a detailed overview of the role of pericytes in inner ear disorders, highlighting the anatomy and physiology in the microvasculature, and analyzing the mechanisms that contribute to the development of the disorders. Furthermore, we explore the potential pericyte-targeted therapies, including antioxidant, anti-inflammatory, and angiogenic approaches, as well as gene therapy strategies.

Utilizing Adenovirus Knob Proteins as Carriers in Cancer Gene Therapy Amidst the Presence of Anti-Knob Antibodies.

Numerous gene therapy drugs for cancer have received global approval, yet their efficacy against solid tumors remains inadequate. Our previous research indicated that the fiber protein, a component of the adenovirus capsid, can propagate from infected cells to neighboring cells that express the adenovirus receptor. We hypothesize that merging this fiber protein with an anti-cancer protein could enable the anti-cancer protein to disseminate around the transfected cells, presenting a novel approach to cancer gene therapy. In our study, we discovered that the knob region of the adenovirus type 5 fiber protein is the smallest unit capable of spreading to adjacent cells in a receptor-specific manner. We also showed that the recombinant knob protein infiltrates cells after dispersing to surrounding cells. To assess the potential of the knob protein to augment gene therapy for solid tumors in mice, we expressed a fusion gene of the A subunit of cytotoxic cholera toxin and the knob region in mouse tumors. We found that this fusion protein only inhibited tumor growth in receptor-expressing mouse melanomas, and this inhibitory effect persisted even in mice with anti-knob antibodies. Our study's findings propose a novel cancer gene therapy strategy that enhances therapeutic effects by specifically delivering therapeutic proteins, expressed from in vivo administered genes, to target molecules. This outcome offers a fresh perspective on gene therapy for solid cancers, and we anticipate that knob proteins will serve as a platform for this method.

Immune Modulation Strategies in Gene Therapy: Overcoming Immune Barriers and Enhancing Efficacy.

The immune system presents significant obstacles to gene therapy, which has limited its use in treating many illnesses. New approaches are needed to overcome these problems and improve the effectiveness of gene therapy. This study explores several techniques to immune regulation within gene therapy, a cutting-edge discipline that aims to optimise results by fine-tuning the immune response. We cover new ways to control the immune system and deliver therapeutic genes just where they are needed, including influencing immunological checkpoints, causing immunotolerance, and making smart use of immunomodulatory drugs. In addition, the study provides insight into new developments in the design of less immunogenic gene delivery vectors, which allow for the extension of transgene expression with minimal adverse immune reactions. In order to maximise the efficacy of gene-based therapies, this review analyses these novel approaches and gives a thorough overview of the present state of the art by addressing obstacles and pointing the way toward future developments in immune regulation. Not only does their integration provide new opportunities for the creation of safer and more effective gene treatments, but it also contains the key to overcome current obstacles.

Preventing and treating neurotrophic keratopathy by a single intrastromal injection of AAV-mediated gene therapy

PurposeNeurotrophic keratopathy (NK) is a degenerative corneal condition resulting from corneal nerve injury. Current therapies, including the recombinant human nerve growth factor (rhNGF) therapy, requires continuous administration. This study aims to develop a novel and highly effective gene therapy strategy for the prevention and treatment of NK. MethodsAdeno-associated virus (AAV) was transduced into corneal stromal cells by intrastromal injection. Three dimensional corneal wholemount imaging with co-immunostaining of ZO-1 and tubulin was utilized to assess the transduction of AAV.rh10. The efficacy of prevention and treatment of NK by a single intrastromal injection of AAV-Ngf was tested using capsaicin mouse model, herpes simplex keratitis (HSK) model, type Ⅱ diabetes model and alkali burn model. rhNGF eye drops served as the positive control. ResultsIntrastromal injection of AAV.rh10 efficiently transduced the subepithelial nerve plexus and retrogradely transported to the trigeminal ganglion (TG). A single injection of AAV.rh10-Ngf can significantly promote corneal nerve repair, accelerate corneal epithelial repair, reduce corneal stromal edema, and improve corneal sensitivity across the four NK models. The therapeutic effects were consistent with those achieved by continuous administration of rhNGF drops by 6 times daily. ConclusionsThis proof-of-concept study demonstrates that AAV.rh10-Ngf gene therapy is a promising method for preventing and treating of NK. Our results underline the potential for developing clinical trials to further explore the safety and efficacy of such gene therapy.

NF-κB-activated oncogene inhibition strategy for cancer gene therapy

NF-κB is a promising target for cancer treatment because of its overactivation in almost all cancers but countless NF-κB inhibitors rarely became clinical drugs due to side effects. In contrast to traditional cancer treatments aimed at inhibiting NF-κB activity, this study develop a novel approach termed HOPE, which focuses on activating the exogenous effector gene CRISPR-Cas13a within cancer cells, achieved by utilizing the NF-κB-specific promoter DMP previously constructed, then targets and suppresses the expression of oncogenes TERT, PLK1, KRAS and MYC at mRNA level. We evaluated the antitumour effects of HOPE in various cultured cells and confirmed it could induce obvious the death of cancer cells without affecting normal cells. By packaging HOPE into adeno-associated virus (AAV) and intravenously injected it to treat mice that were subcutaneously transplanted with colorectal cancer. This validated that rAAV-HOPE could significantly inhibit tumour growth without side effects. Based on the scRNA-seq data, we observed that HOPE could activate the immune system and decrease the proportion of cancer cells, particularly reducing the stemness of cancer cells. This study elucidates an important role of HOPE in inhibiting cancer cell growth both in vitro and in vivo, additionally provides a novel therapeutic technology for cancer gene therapy.

Insights into multidrug resistance mechanisms: Exploring distinct miRNAs as prospective therapeutic agents in triple negative breast cancer

Triple Negative Breast Cancer (TNBC) constitutes 12–17 % of breast cancers and is distinguished by the absence of hormone receptor expression, deviating from other breast cancer types. Coupled with its elevated proliferation index, TNBC develops multidrug resistance, diminishing treatment efficacy, weakening disease prognosis, and leading to an aggressive clinical course. This study aimed to assess ABCB1, ABCC1, ABCG2, and ABCC9 gene expression, which play a primary role in the development of multidrug resistance, alongside miRNAs (miR-466, miR-4539, miR-659-3p, miR-3123, miR-3133, and miR-655-3p) targeting these genes. While ABCB1 (p = 0.433), ABCC1 (p < 0.05), and ABCG2 (p < 0.05) exhibited increased expression in tumor tissues, ABCC9 (p = 0.587) did not. miR-466 (p = 0.802), miR-4539 (p = 0.732), miR-659-3p (p = 0.807), and miR-3123 (p = 0.980) were upregulated, whereas miR-3133 (p < 0.05) and miR-655-3p (p = 0.190) were downregulated. Within the scope of our study, we also evaluated the clinical parameters like tumor size, stage, and neoadjuvant treatment that significantly impacted Progression Free Survival (PFS) and Overall Survival (OS). Considering these results, we found that the metastatic status significantly influenced PFS and OS. Chemotherapeutics were found to not affect survival times. Assessing the impact of miRNAs, which we view as potential therapeutic targets, on average survival revealed that elevated miR-3133 expression was correlated with shorter PFS and OS, whereas decreased miR-655-3p expression was associated with longer PFS and OS. In summary, the relevant miRNAs could serve as predictive biomarkers for drug response and aid in developing miRNA-targeted gene therapy strategies.