Structural characterization and activation of nature’s fuels of life

Hydrolysis of nucleotide triphosphates (NTPs) is known to drive highly important cellular processes. Of these NTPs, adenine triphosphate (ATP) is mainly studied as it is concerned to be the central fuel of living cells. Gas-phase studies have been performed to elucidate the conformational preference of NTPs focusing on the dependence of the nucleobase and extent of deprotonation. Therefore, a combination of infrared (IR) spectroscopy and mass spectrometry has been applied to ATP, CTP and GTP. IR action spectroscopy and supporting theoretical studies showed that the attached nucleobase has little influence on the three-dimensional structure of the phosphate tail. The favorable deprotonation sites of the singly and doubly deprotonated NTPs have been assigned. This can lead to a better explanation of the behavior and selection of NTPs in living cells

Tyrosine kinase inhibitor resistance in de novo BCR::ABL1–positive BCP-ALL beyond kinase domain mutations

A better understanding of ABL1 kinase domain mutation–independent causes of tyrosine kinase inhibitor (TKI) resistance is needed for BCR::ABL1–positive B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Although TKIs have dramatically improved outcomes, a subset of patients still experiences relapsed or refractory disease. We aimed to identify potential biomarkers of intrinsic TKI resistance at diagnosis in samples from 32 pediatric and 19 adult patients with BCR::ABL1–positive BCP-ALL. Reduced ex vivo imatinib sensitivity was observed in cells derived from newly diagnosed patients who relapsed after combined TKI and chemotherapy treatment compared with cells derived from patients who remained in continuous complete remission. We observed that ex vivo imatinib resistance was inversely correlated with the amount of (phosphorylated) BCR::ABL1/ABL1 protein present in samples that were taken at diagnosis without prior TKI exposure. This suggests an intrinsic cause of TKI resistance that is independent of functional BCR::ABL1 signaling. Simultaneous deletions of IKZF1 and CDKN2A/B and/or PAX5 (IKZF1plus), as well as deletions of PAX5 alone, were related to ex vivo imatinib resistance. In addition, somatic lesions involving ZEB2, SETD2, SH2B3, and CRLF2 were associated with reduced ex vivo imatinib sensitivity. Our data suggest that the poor prognostic value of IKZF1(plus) deletions is linked to intrinsic mechanisms of TKI resistance other than ABL1 kinase domain mutations in newly diagnosed pediatric and adult BCR::ABL1–positive BCP-ALL.</p