DNA-PK and ATM inhibition strongly radiosensitized uveal melanoma models to proton therapy

Blocking DNA double-strand-break repair enhanced proton response across seven uveal melanoma spheroid models, with DNA-PK inhibition showing the strongest effect.

KEY POINTS

  • Investigators studied seven 3D uveal melanoma spheroid models—92.1, Mel202, Mel270, MP41, MP46, OMM1 and OMM2.3—and combined X-rays or proton beam therapy with inhibitors targeting three major DNA double-strand-break response kinases: ATM, ATR and DNA-PK.
  • Baseline radiosensitivity varied substantially. After 2 Gy of X-rays, growth of the 92.1 model fell by approximately 60%, whereas MP41 and OMM2.3 showed only around 10–20% reduction, providing a useful range of intrinsically sensitive and resistant models.
  • With X-rays, DNA-PK inhibition produced the largest radiosensitization, with dose-enhancement ratios of 2.5–10.0, followed by ATM inhibition at 1.6–8.0 and ATR inhibition at 1.3–5.6. The magnitude depended heavily on the intrinsic radiosensitivity of each cell line.
  • The effect persisted with protons. Across proton-treated spheroids, DNA-PK inhibition produced dose-enhancement ratios of approximately 1.7–5.6 and ATM inhibition 1.9–5.6, again making these the strongest of the tested repair targets.
  • Mechanistic experiments showed that the inhibitors prevented normal resolution of radiation-induced DNA double-strand breaks. After proton therapy, DNA-PK inhibition almost completely prevented resolution of γH2AX and 53BP1 signaling through 24 hours, while ATM and ATR inhibition also prolonged damage signaling into the 24–48-hour period.
  • X-ray combination treatment also increased micronuclei and other chromosomal abnormalities. ATM and DNA-PK inhibition prolonged G2/M arrest at 24 hours, consistent with persistent unrepaired damage rather than simply nonspecific suppression of cell growth.
  • Some enhancement ratios were actually lower with protons than with X-rays, partly because proton irradiation alone already produced greater growth suppression in several models. The experimental proton beam also differed geometrically and had a moderately higher reported linear energy transfer, complicating direct modality comparisons.

CLINICAL TAKEAWAY

DNA-repair inhibition offers a biologically plausible route to increasing uveal-melanoma radiosensitivity and, in principle, could eventually permit better control or lower ocular dose. But these experiments stop at 3D tumor spheroids: there is no evidence yet that systemic or local ATM/DNA-PK inhibition can be delivered safely enough to improve the therapeutic ratio in an eye, where protection of retina, optic nerve and other normal tissues is the entire point.

SOURCE

Frontiers in Oncology