A pilot study of the use of dynamic analysis of cell-free DNA from aqueous humor and vitreous fluid for the diagnosis and treatment monitoring of vitreoretinal lymphomas

Abstract

The diagnosis of vitreoretinal lymphoma (VRL), a rare subtype of primary central nervous system lymphoma (PCNSL), is challenging. We aimed to investigate the mutational landscape of VRL by sequencing circulating tumor DNA (ctDNA) from aqueous humor (AH) and/or vitreous fluid (VF), as well as the application of ctDNA sequencing to diagnosis and treatment monitoring. Baseline AH and/or VF specimens from 15 VRL patients underwent comprehensive genomic profiling using targeted next-generation sequencing. The molecular profiles of paired baseline AH and VF specimens were highly concordant, with comparable allele frequencies (AFs). However, the genetic alterations detected in cerebrospinal fluid (CSF) ctDNA only partially overlapped with those from simultaneously-collected AH/VF samples, with much lower AFs. Serial post-treatment AH or VF samples were available for five patients and their ctDNA AF changes displayed a similar trend as the changes in interleukin 10 (IL-10) levels; an indicator of treatment responses. A cohort of 23 PCNSL patients was included as a comparison group for the genetic landscape and evaluations of the efficacy of ibrutinib. More MYD88 mutations, but fewer IRF4 mutations and CDKN2A/B copy number losses were observed in the baseline samples of PCNSL than VRL patients. The objective response rate of ibrutinib treatment was much higher for PCNSL patients (64.7%, 11/17) than VRL (14.3%, 1/7) patients. In summary, we provided valuable clinical evidence that AH is a good source of tumor genomic information and can be substituted for VF. Moreover, molecular profiling of AH has clinical utility for the diagnosis of VRL and treatment monitoring.

Circulating cell-free DNA and IL-10 from cerebrospinal fluids aid primary vitreoretinal lymphoma diagnosis

“…Several studies demonstrated the presence of MYD88 L265P mutation in the aqueous humor and vitreous fluid of vitreoretinal lymphoma patients. In different PVRL cohorts, the reported sensitivity of MYD88 L265P mutation detection ranged from 25% to 88.9%, with direct Sanger sequencing of polymerase chain reaction (PCR), droplet digital PCR, or sequencing (12)(13)(14)(15)(16)(17)(18)(19). The vitreous fluid samples showed higher positive rate than paired aqueous humor samples (14).…”

“…In the recent years, numerous studies have focused on exploring potential ancillary techniques and biomarkers for primary vitreoretinal lymphoma (PVRL) diagnosis. The detection of PVRL characteristic gene mutations, including MYD88 L265P and CD79B , and immunoglobulin gene rearrangement ( 4 , 12 – 21 ) from aqueous humor or vitreous fluid has been considered effective diagnostic approaches. Meanwhile, elevate aqueous humor or vitreous fluid interleukin-10 (IL-10) levels and elevated IL-10/IL-6 ratios have been considered as useful biomarkers ( 22 ).…”

Nupr1 Negatively Regulates Endothelial to Hematopoietic Transition in the Aorta‐Gonad‐Mesonephros Region

“…Considering the potential role of Nupr1 as a negative regulator of the cell cycle and iron‐induced cell death (ferroptosis), [ 29,31 ] we first examined the cell cycle states of different EHT‐related populations at E10.0/E11.0, including immunophenotypic AECs, HECs and IAHC cells, by Hoechst and Ki67 staining. Notably, no difference in cell cycle status between control and cKO embryos was detected (Figure 2E and Figure S5A, Supporting Information).…”

“…Notably, a recent study revealed that Nupr1, a stress‐responsive molecule involved in multiple biological contexts, is an adult HSC quiescence regulator that coordinates with the p53 signaling pathway. [ 29 ] However, the physiological function of Nupr1 in blood generation during embryogenesis remains unknown.…”

Diagnostic value of genetic mutation analysis and mutation profiling of cell‐free DNA in intraocular fluid for vitreoretinal lymphoma

“…The 5 most frequently mutated genes found in these patients were proviral integration site for moloney murine leukemia virus-1 (PIM1) in 90.9%; myeloid differentiation primary response protein 88 (MyD88) in 77.3%; CD79B in 50.0%; ETS variant 6 (ETV6) in 50.0%; and interferon regulatory factor 4 (IRF4) in 50.0% (Figure 1G). More PIM1 and IRF4 mutations, but fewer MyD88 mutation were observed in VRL patients than primary central nervous system lymphomas (PCNSL) patients as we previously reported [6]. Although two patients with uveitis were ctDNA-positive, their ctDNA concentrations were low; the observed mutations in the two patients with ctDNA-positive uveitis are uncommon in B-cell lymphomas, and we considered them to be clonal hematopoietic mutations (Supplementary Table S6).…”