Validation and Early Commercialization of the ENVISAGE Assay, a Prognostic Test for Barrett's Esophagus

Barrett食管（BE）是食管高血压的最强已知风险因素，也是食管高血压的专性前体， 分级异型增生（HGD）和腺癌（EAC）。然而，确定未来肿瘤进展风险 在BE患者中是相当具有挑战性的：目前的风险估计仅基于高度可变的，主观的， 非依赖性组织病理学诊断（即，非异型增生（ND），低度异型增生（LGD）， 不确定的异型增生（IFD）或高度异型增生（HGD）。我们的ENVISAGE检测是一种经过验证的 (CLIA#21D2256153）基于4基因加患者年龄DNA的实验室开发测试（LDT） 在Capsulomics开发的基于甲基化的PCR检测方法，基于进行的基础研究 约翰霍普金斯大学（JHU）。我们的初步研究产生了一个准备推出的第一代检测 根据分子水平准确预测BE患者未来肿瘤进展的风险， 生物标志物。这种预后分析对患者进行风险分层，大大改善了临床决策， 灵敏度为71%，特异性为90%，优于任何其他临床可用的测试。 尽管取得了初步的成功，我们已经确定需要进一步改进，并制定了一个强有力的 商业化道路。有了这项直接进入第二阶段的建议的资金，我们将大大提高 改进以生成更灵敏、更稳健验证的第二代产品，同时 为当前和未来的战略商业化和市场采用奠定基础 测定。目标1将解决由于使用有限数量、低质量的片段化 通过设计一种基于多重PCR的新检测方法从活检标本中提取DNA。这一进步将 还提高了吞吐量、效率、成本和处理速度。目标2和3将加强我们的研究 人口，同时纳入新的多重协议，从目标1，保留和完善我们的算法， 产生第二代ENVISAGE检测，并更清晰地描述哪些患者将落入 改进临床管理的"中等"风险类别。Aim 4将建立早期商业化 同时为未来推出的第二代检测开发平台 产品因此，这一直接进入II期SBIR的提议将扩展、完善和验证ENVISAGE测定， 一种新的LDT，用于对BE患者未来的肿瘤进展风险进行分层，从而改善临床 决策的

Barrett’s esophagus (BE) is the strongest known risk factor for, and obligate precursor of, esophageal high- grade dysplasia (HGD) and adenocarcinoma (EAC). However, determining future neoplastic progression risk is quite challenging in BE patients: current risk estimation is based solely on highly variable, subjective, observer-dependent histopathologic diagnoses (i.e., non-dysplastic (ND), Low-Grade Dysplasia (LGD), Indeterminate for Dysplasia (IFD), or High-Grade Dysplasia (HGD). Our ENVISAGE assay is a validated (CLIA # 21D2256153) laboratory-developed test (LDT) based on a 4-gene-plus-patient age DNA methylation-based PCR assay that was developed at Capsulomics built on foundational studies performed at Johns Hopkins University (JHU). Our preliminary studies yielded a ready-to-launch 1st-generation assay that accurately predicts the risk of future neoplastic progression in BE patients based on levels of molecular biomarkers. This prognostic assay risk-stratifies patients to vastly improve clinical decision-making, with a sensitivity of 71% and a specificity of 90%, outperforming any other clinically available tests. Notwithstanding this early success, we have identified a need for further improvement as well as a robust commercialization pathway. With funding of this direct-to-phase II proposal, we will make significant improvements to generate a more sensitive, more robustly validated second-generation product, while building a foundation for strategic commercialization and market adoption of both the current and future assays. Aim 1 will address assay sensitivity challenges due to using limited-quantity, low-quality fragmented DNA from biopsy specimens by engineering a new assay based on multiplex PCR. This advancement will also improve throughput, efficiency, cost, and processing speed. Aims 2 and 3 will augment our study population while incorporating the new multiplex protocol from Aim 1, retain and refine our algorithm to produce a 2nd-generation ENVISAGE assay, and more sharply delineate which patients will fall into the “Intermediate” risk category for improved clinical management. Aim 4 will establish early commercialization of the 1st-generation assay as an LDT while developing a platform for the future launch of our 2nd-generation product. Thus, this direct-to-phase II SBIR proposal will expand, refine, and validate the ENVISAGE assay, a novel LDT for stratifying future neoplastic progression risk in BE patients, thereby improving clinical decision-making.