如果我们可以通过单一方法来治疗甚至治愈疾病,该有多好?基因治疗的愿景,是可以纠正疾病的潜在遗传原因。尽管基因疗法具有令人兴奋的潜力,但由于安全性和有效性方面的问题,以及早期临床试验的失败强调需要进一步深入了解基因疗法的基础科学。欢迎了解 NanoTemper 是如何帮助研究人员开发更安全,更有效的基因疗法。

安全性和有效性推动技术前行

病毒载体腺相关病毒(AAV)和慢病毒,以及基因编辑技术是开发治疗多种疾病(例如镰状细胞性贫血,亨廷顿氏病和某些类型的癌症)的关键工具。为了达到功效,这些疗法必须能高效且安全地将核酸递送至靶细胞。精准表征病毒载体或基因编辑蛋白与靶细胞之间的相互作用是提高治疗安全性和有效性的一种高效方法。

Learn how knowing binding affinity can help you develop safer and more potent gene therapies

 

观看在线讲座

病毒载体

AAV 和慢病毒被用作病毒载体,可以将修饰的基因递送至靶细胞。研究人员致力于开发出仅针对靶向特定细胞的更好的载体,同时避免触发患者的免疫反应,并且可以有效地持续进行治疗。上述目标可以通过表征分子相互作用并根据其热变性数据对病毒载体进行分析来实现。

 


 

提高 CD19 CAR-T 细胞疗法的有效性

CD19 CAR-T cell therapy has shown promising results in treating B-cell malignancies using murine CD19 CAR — but can lead to immune recognition in some patients and make the treatment ineffective. This study sought to find out whether humanized CD19 CAR would resolve this problem. MST was used to measure the affinity between CD19 and the murine or humanized CD19 CAR. The humanized CD19 CAR had a 6-fold greater affinity for its human target which improved the outcome of the therapy when used to treat patients — a higher number of therapeutic T cells with increased anti-tumor activity.

了解更多

 


 

在任何缓冲液中鉴定 AAV 血清型

AAV tropism – which determines the target cell or tissue – differs between AAV serotypes. The thermal stability of the AAV capsid is one parameter that can be used to differentiate between serotypes. In this study, nanoDSF was used to measure the thermal stability of different AAV capsids during the development and production process of AAVs. nanoDSF allowed researchers to rapidly identify AAV serotypes in many different formulation and storage buffers.

了解更多

 


 

See how changes in production method affect your AAV serotypes

Scale-up of recombinant AAVs is key for manufacturers aiming to get their gene therapies to market. While preclinical AAVs are produced in human-derived HEK293 cells, these do not offer ideal throughput for scale up. This study investigated whether the baculovirus-sf9 system made an appropriate substitute to HEK293s, and in particular whether the post-translational modifications (PTMs) of the insect-cell system were sufficient to prevent an adverse reaction in patients. The group used nanoDSF to characterize the differences between different capsid serotypes, as well as different modes of production for the same serotype.

了解更多

Discover solutions for analytical characterization during AAV vector development

 

观看在线讲座

Payload delivery

In order to design a good gene therapy vector, researchers must consider not only capsid formation and structure, but also how well it delivers the genetic payload to the cell. Gene therapy optimization requires understanding how much genetic material is loaded into the vector, as well as the mechanism of payload release. Avoiding the loss of DNA or RNA during storage and transport is also a critical concern for those working in scale-up and manufacturing of gene therapy products.

 


 

Monitor vector stability and prevent DNA loss in AAVs

Manufacturing a successful gene therapy vector relies on the stability of its capsid. This means not only ensuring the capsids are serotyped properly, but also that none of the genetic material is lost over the course of the manufacturing process. This group used nanoDSF to probe vector stability and relate it to the amount of DNA lost in different storage conditions.

了解更多

Learn about infectious viral mechanisms, which can improve our understanding of AAVs

 

See virology spotlight

基因编辑技术

基因编辑技术可以通过有针对性地对体内基因组进行编辑(包括基因添加,删除和校正)来提供基因治疗。尤其是,CRISPR-Cas9 体系为治疗遗传性疾病提供了令人兴奋的新的可能性。现在,研究人员正在努力通过使用 Cas9 的变体来消除脱靶编辑或寻找可替代的递送系统来使基因编辑工具变得更安全可靠。

 


 

使用其他 Cas9 变体提高基因编辑特异性

Gene editing with CRISPR-Cas9 has shown great promise in the treatment of diseases such as sickle cell anemia. However, off-target gene editing has been reported for some Cas9 proteins, like SpCas9. This study used MST to show that a different variant of Cas9, FnCas9, has a higher specificity for its intended target and low off-target binding. FnCas9 was then used to successfully correct sickle cell mutations in the patient-derived pluripotent stem cells.

了解更多

 


 

设计特异性更强的核酸输送系统

The use of viral vectors still raises concerns about safety due to off-target delivery of nucleic acids. To improve target specificity, the authors designed a chromatin-based nucleic acid delivery system that incorporates antibodies specific to cell surface elements. The key to the success of this system is the efficient capture of the antibody to the chromatin, and MST was used to quantify the interaction between different antibodies and the chromatin. This allowed them to find the best antibody for constructing a highly efficient and specific chromatin-based delivery system for CRISPR-Cas9 gene editing.

了解更多

Want to see more gene therapy publications?

 See resources

汉语日本語English