Exploring Biochemical Characteristics of Pediatric Hyperdiploid Acute Lymphoblastic Leukemia by Raman Spectroscopy

<i>Hyperdiploid</i> (HD) B-cell acute lymphoblastic leukemia (ALL) is widely recognized as the most common molecular subtype of leukemia, characterized by the presence of supernumerary chromosomes in the leukemic karyotype. While HD B-ALL is often associated with a favorable prognosis, an important subset of patients still experience relapse, reflecting the biological heterogeneity of this subtype. Current genomic and epigenetic research has shed light on the molecular complexity of HD B-ALL, yet rapid methods for capturing both the metabolic state and the chromosomal content of individual cells remain limited. Here, we introduce a novel Raman spectroscopy (RS)-based approach for the single-cell analysis of HD B-ALL. By detecting characteristic spectroscopic signatures of nucleic acids, proteins, and lipids, RS not only distinguishes malignant cells from normal B cells, but also discriminates between HD B-ALL and other molecular subtypes, including <i>TCF3-PBX1</i>, <i>KMT2A-r</i>, <i>BCR-ABL1</i>, and <i>TEL-AML1</i>. Notably, we developed a partial least-squares regression (PLS-R) model capable of accurately predicting chromosome number from each cell's Raman spectrum, thereby linking molecular fingerprints directly to genomic aberrations. This integrative spectroscopic strategy captures disease heterogeneity and informs therapeutic strategies. Taken together, our proof-of-concept findings highlight RS as a powerful, noninvasive tool for quantifying chromosomal alterations and metabolic phenotypes, adding crucial insights into the complex biology of HD B-ALL and paving the way for broader applications in precision medicine.