Abstract

Acute leukemias (ALs) are often associated with chromosomal translocations, in particular, KMT2A/MLL gene rearrangements. Identification or confirmation of these translocations is carried out by a number of genetic and molecular methods, some of which are routinely used in clinical practice, while others are primarily used for research purposes. In the clinic, these methods serve to clarify diagnoses and monitor the course of disease and therapy. On the other hand, the identification of new translocations and the confirmation of known translocations are of key importance in the study of disease mechanisms and further molecular classification. There are multiple methods for the detection of rearrangements that differ in their principle of operation, the type of problem being solved, and the cost-result ratio. This review is intended to help researchers and clinicians studying AL and related chromosomal translocations to navigate this variety of methods. All methods considered in the review are grouped by their principle of action and include karyotyping, fluorescence in situ hybridization (FISH) with probes for whole chromosomes or individual loci, PCR and reverse transcription-based methods, and high-throughput sequencing. Another characteristic of the described methods is the type of problem being solved. This can be the discovery of new rearrangements, the determination of unknown partner genes participating in the rearrangement, or the confirmation of the proposed rearrangement between the two genes. We consider the specifics of the application, the basic principle of each method, and its pros and cons. To illustrate the application, examples of studying the rearrangements of the KMT2A/MLL gene, one of the genes that are often rearranged in AL, are mentioned.

摘要

急性白血病 (AL) 通常与染色体易位有关，尤其是KMT2A/MLL基因重排。这些易位的鉴定或确认是通过许多遗传和分子方法进行的，其中一些在临床实践中常规使用，而另一些则主要用于研究目的。在临床中，这些方法用于澄清诊断并监测疾病和治疗过程。另一方面，新易位的鉴定和已知易位的确认在疾病机制研究和进一步的分子分类中至关重要。有多种检测重排的方法，它们的操作原理、解决问题的类型和成本-结果比都不同。本综述旨在帮助研究 AL 和相关染色体易位的研究人员和临床医生使用各种方法。审查中考虑的所有方法均按其作用原理分组，包括核型分析、荧光原位杂交 (FISH) 探针检测整个染色体或单个基因座、基于 PCR 和逆转录的方法以及高通量测序。所描述方法的另一个特征是要解决的问题类型。这可以是新重排的发现、参与重排的未知伙伴基因的确定，或两个基因之间提议的重排的确认。我们考虑了应用程序的具体情况、每种方法的基本原理及其优缺点。为了说明应用程序，提到了KMT2A/MLL基因，AL中经常重排的基因之一。