Constitutional mismatch repair deficiency (CMMRD) is a highly penetrant cancer predisposition syndrome caused by biallelic mutations in mismatch repair (MMR) genes. As several cancer syndromes are clinically similar, accurate diagnosis is critical to cancer screening and treatment. As genetic diagnosis is confounded by 15 or more pseudogenes and variants of uncertain significance, a robust diagnostic assay is urgently needed. We sought to determine whether an assay that directly measures MMR activity could accurately diagnose CMMRD.In vitro MMR activity was quantified using a 3'-nicked G-T mismatched DNA substrate, which requires MSH2-MSH6 and MLH1-PMS2 for repair. We quantified MMR activity from 20 Epstein-Barr virus-transformed lymphoblastoid cell lines from patients with confirmed CMMRD. We also tested 20 lymphoblastoid cell lines from patients who were suspected for CMMRD. We also characterized MMR activity from patients with neurofibromatosis type 1, Li-Fraumeni syndrome, polymerase proofreading-associated cancer syndrome, and Lynch syndrome.All CMMRD cell lines had low MMR activity (n = 20; mean, 4.14 ± 1.56%) relative to controls (n = 6; mean, 44.00 ± 8.65%; P < .001). Repair was restored by complementation with the missing protein, which confirmed MMR deficiency. All cases of patients with suspected CMMRD were accurately diagnosed. Individuals with Lynch syndrome (n = 28), neurofibromatosis type 1 (n = 5), Li-Fraumeni syndrome (n = 5), and polymerase proofreading-associated cancer syndrome (n = 3) had MMR activity that was comparable to controls. To accelerate testing, we measured MMR activity directly from fresh lymphocytes, which yielded results in 8 days.On the basis of the current data set, the in vitro G-T repair assay was able to diagnose CMMRD with 100% specificity and sensitivity. Rapid diagnosis before surgery in non-neoplastic tissues could speed proper therapeutic management.
ABSTRACT Although right ventricle (RV) dysfunction drives clinical worsening in pulmonary hypertension (PH), information about RV function has not been well integrated in PH risk assessment. The gold standard for assessing RV function and ventriculo‐arterial coupling is the construction of multi‐beat pressure–volume (PV) loops. PV loops are technically challenging to acquire and not feasible for routine clinical use. Therefore, we aimed to map standard clinically available measurements to emergent PV loop phenotypes. One hundred and one patients with suspected PH underwent right heart catheterization (RHC) with exercise, multi‐beat PV loop measurement, and same‐day cardiac magnetic resonance imaging (CMR). We applied unsupervised k ‐means clustering on 10 PV loop metrics to obtain three patient groups with unique RV functional phenotypes and times to clinical worsening. We integrated RHC and CMR measurements to train a random forest classifier that predicts the PV loop patient group with high discrimination (AUC = 0.93). The most informative variable for PV loop phenotype prediction was exercise mean pulmonary arterial pressure (mPAP). Distinct and clinically meaningful PV loop phenotypes exist that can be predicted using clinically accessible hemodynamic and RV‐centric measurements. Exercise mPAP may inform RV pressure–volume relationships.
