Laboratorio de Genética y Cáncer, Instituto Nacional de Pediatría, Mexico City 04530, Mexico.
Cátedra CONACYT-Instituto Nacional de Pediatría, Mexico City 04530, Mexico.
Int J Mol Sci. 2022 Aug 24;23(17):9587. doi: 10.3390/ijms23179587.
Ph-like subtypes with abnormalities are frequent among Hispano-Latino children with pre-B ALL. Therefore, there is solid ground to suggest that this subtype is frequent in Mexican patients. The genomic complexity of Ph-like subtype constitutes a challenge for diagnosis, as it requires diverse genomic methodologies that are not widely available in diagnostic centers in Mexico. Here, we propose a diagnostic strategy for Ph-like ALL in accordance with our local capacity. Pre-B ALL patients without recurrent gene fusions (104) were classified using a gene-expression profile based on Ph-like signature genes analyzed by qRT-PCR. The expressions of the transcript and protein were determined by qRT-PCR and flow cytometry. The , rearrangements, and Ik6 isoform were screened using RT-PCR and FISH. Surrogate markers of Jak2-Stat5/Abl/Ras pathways were analyzed by phosphoflow. Mutations in relevant kinases/transcription factors genes in Ph-like were assessed by target-specific NGS. A total of 40 patients (38.5%) were classified as Ph-like; of these, 36 had abnormalities associated with Jak2-Stat5 and 4 had Abl. The rearrangements , and iAMP21 were particularly frequent. We propose a strategy for the detection of Ph-like patients, by analyzing the overexpression/genetic lesions of , the Abl phosphorylation of surrogate markers confirmed by gene rearrangements, and Sanger sequencing.
在有 Pre-B ALL 的西班牙裔拉丁裔儿童中，存在异常的 Ph-like 亚型很常见。因此，有充分的理由表明这种亚型在墨西哥患者中很常见。Ph-like 亚型的基因组复杂性给诊断带来了挑战，因为它需要多样化的基因组方法，但这些方法在墨西哥的诊断中心并不广泛。在这里，我们根据当地的能力提出了 Ph-like ALL 的诊断策略。使用基于 qRT-PCR 分析的 Ph-like 特征基因的基因表达谱对无复发性基因融合的 Pre-B ALL 患者（104 例）进行分类。通过 qRT-PCR 和流式细胞术确定转录物和蛋白质的表达。使用 RT-PCR 和 FISH 筛选 、 重排和 Ik6 同工型。通过磷酸化流式细胞术分析 Jak2-Stat5/Abl/Ras 途径的替代标志物。通过靶向特异性 NGS 评估 Ph-like 中相关激酶/转录因子基因的突变。共有 40 例（38.5%）患者被归类为 Ph-like；其中 36 例存在与 Jak2-Stat5 相关的异常，4 例存在 Abl。重排 、 和 iAMP21 特别常见。我们提出了一种检测 Ph-like 患者的策略，通过分析 、 Abl 磷酸化的替代标志物的过度表达/遗传病变进行检测，这些替代标志物通过基因重排和 Sanger 测序得到确认。
