Synthetic introns for selective targeting of RNA splicing factor-mutant leukemia

用于选择性靶向RNA剪接因子突变型白血病的合成内含子

基本信息

批准号：
10722782
负责人：
Omar Abdel-Wahab
金额：
$ 74.86万
依托单位：
FRED HUTCHINSON CANCER CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10722782
关键词：
3&apos Splice Site Acute Myelocytic Leukemia Address Affect Alleles Anatomy Biomedical Engineering Biotechnology Breast Cancer Cell Cancer Biology Cancer Patient Cells Chronic Myelomonocytic Leukemia Clinical Disease Dissection Drug Delivery Systems Dysmyelopoietic Syndromes Elements Engineering Event Exons FDA approved Frequencies Ganciclovir Gene Expression Growth Health Hematologic Neoplasms Hematological Disease Hematopoietic Hematopoietic Neoplasms In Vitro Individual Interleukin-15 Introns Left Lesion Leukemic Cell Malignant Neoplasms Mediating Melanoma Cell Messenger RNA Methods Molecular Mutate Mutation Myeloid Leukemia Myeloproliferative disease Nature Patient Care Patients Pattern Pharmaceutical Preparations Physicians Precision therapeutics Production Proteins RNA Splicing Recurrence Refractory SRSF2 gene Safety Scientist Simplexvirus Solid Neoplasm Spliceosomes System Technology Testing Therapeutic Thymidine Kinase Tissues Work Xenograft procedure cancer cell cancer type cell killing cell type clinical phenotype conventional therapy efficacy evaluation experimental study functional genomics immunostimulatory therapy in vivo insight leukemia lipid nanoparticle loss of function malignant breast neoplasm melanoma mutant neoplasm relapse new technology novel therapeutics protein expression selective expression side effect suicide gene therapeutic protein therapeutic target tumor tumorigenic

项目摘要

SUMMARY Many cancers carry recurrent, change-of-function mutations affecting RNA splicing factors, resulting in sequence-specific changes in RNA splicing that promote disease initiation and progression. These “spliceosomal mutations” are the most common class of mutations in myelodysplastic syndromes (MDS) and related hematologic disorders, which have few effective, FDA-approved treatments. Despite the high frequency of spliceosomal mutations and corresponding need for new therapeutics, there currently exist no therapies that specifically and selectively target these lesions. Here, we propose to address this clinical need by creating new precision therapeutics that selectively kill cells with spliceosomal mutations. Our interdisciplinary team consists of a physician-scientist with expertise in cancer biology and patient care (Abdel-Wahab), a basic scientist with expertise in RNA splicing and functional genomics (Bradley), and a bioengineer with expertise in drug delivery (Heller). In preliminary experiments, we developed the “synthetic intron” technology to harness altered RNA splicing activity caused by spliceosomal mutations to drive cancer-specific gene expression, showed that synthetic introns permit highly selective expression of therapeutic payloads in cancer cells while leaving healthy cells unharmed, and used this system to suppress the growth of diverse cancer types in vivo (North et al, Nature Biotechnology, 2022). We additionally demonstrated that synthetic introns enable simultaneous and selective delivery of multiple therapeutic payloads and allow for detailed mechanistic dissection of the cis- and trans-acting sequence elements and splicing factors that govern pro-tumorigenic mis-splicing caused by recurrent spliceosomal mutations. We will now build on these preliminary studies to develop synthetic intron-based therapeutics for myeloid neoplasms, including MDS, acute myeloid leukemia (AML), and chronic myelomonocytic leukemia (CMML), and additionally utilize synthetic introns to understand the mechanistic basis for aberrant splicing in these diseases as follows: Aim 1, Dissect and exploit the molecular mechanisms underlying common as well as allele-specific splicing changes induced by different SF3B1 mutations; Aim 2, Develop synthetic introns that enable selective therapeutic protein expression for each of the commonly mutated RNA splicing factors in leukemia; Aim 3, Optimize in vivo delivery and rigorously test an immunostimulatory therapy for treating SF3B1-mutant hematopoietic malignancies. The significance of these studies is that they will develop a new technology that enables mechanistic studies of cancer-associated spliceosomal mutations and also provides a specific means for therapeutically targeting these mutations. The health relatedness is that the proposed work will create specific therapeutic products for treating cancer types that currently have few effective, FDA- approved treatments.

概括许多癌症携带反复发生的功能改变突变，影响 RNA 剪接因子，从而导致 RNA剪接的序列特异性变化促进疾病的发生和进展。 “剪接体突变”是骨髓增生异常综合征 (MDS) 中最常见的一类突变相关的血液系统疾病，尽管频率很高，但经 FDA 批准的有效治疗方法却很少。由于剪接体突变和相应的新疗法的需要，目前尚无疗法可以专门且选择性地针对这些病变。在这里，我们建议通过创建新的精准疗法来选择性地满足这一临床需求我们的跨学科团队由具有专业知识的医师科学家组成。癌症生物学和患者护理 (Abdel-Wahab) 博士，一位基础科学家，在 RNA 剪接和功能基因组学（Bradley）和具有药物输送专业知识的生物工程师（Heller）处于初步阶段。实验中，我们开发了“合成内含子”技术来利用RNA剪接活性由剪接体突变引起，驱动癌症特异性基因表达，表明合成内含子允许在癌细胞中高度选择性地表达治疗有效负载，同时保持健康细胞未受到伤害，并使用该系统抑制体内多种癌症类型的生长（North 等人，2017）。 al，Nature Biotechnology，2022）我们还证明了合成内含子可以同时实现和实现。选择性递送多种治疗有效负载，并允许对顺式和控制促肿瘤发生错误剪接的反式作用序列元件和剪接因子反复出现的剪接体突变。我们现在将在这些初步研究的基础上开发基于内含子的合成疗法骨髓肿瘤，包括 MDS、急性髓细胞白血病 (AML) 和慢性粒单核细胞白血病 (CMML)，并另外利用合成内含子来了解异常剪接的机制基础这些疾病如下：目标1，剖析和探索常见疾病的分子机制作为不同 SF3B1 突变诱导的等位基因特异性剪接变化；目标 2，开发合成内含子使每个常见突变的 RNA 剪接因子能够选择性表达治疗蛋白目标 3，优化体内递送并严格测试免疫刺激疗法的治疗效果 SF3B1 突变型造血系统恶性肿瘤。这些研究的意义在于他们将开发一种新的该技术能够对癌症相关剪接体突变进行机制研究，并提供针对这些突变进行治疗的具体方法与健康相关性是拟议的工作。将创造特定的治疗产品来治疗目前几乎没有有效的癌症类型，FDA- 批准的治疗方法。