# Analytic and Clinical Validation of a 7-plex MIF Assay for Predictive Response of Advanced NSCLC to Anti-PD-1 based Therapy

> **NIH NIH UH2** · JOHNS HOPKINS UNIVERSITY · 2022 · $224,347

## Abstract

PROJECT SUMMARY
 PD-L1 membranous (cell surface) expression in pretreatment biopsies is the most
commonly used correlate of the likelihood of response to anti-PD-1 therapy. The general finding
of an association of PD-L1 expression with tumor response to anti-PD-1 therapy has been
substantiated across tens of thousands of patients with numerous tumor types treated with anti-
PD-(L)1. However, while PD-L1 expression enriches for response to anti-PD-(L)1, it is not
sufficient. Additionally, while pathologists demonstrate good reproducibility for scoring tumor cell
(TC) PD-L1 expression by chromogenic IHC and light microscopy, they have poor reproducibility
for scoring PD-L1 expression on immune cells (IC).
 Other related features which have been shown to improve on the PD-L1 biomarker include
the proximity of PD-1 to PD-L1, the density of CD8+FoxP3+ cells, and CD68 and tumor marker
immunostains, the latter of which help identify co-expression of PD-L1 on ICs and TCs,
respectively. A quantitative multiplex immunofluorescence assay which captures all of these
features has been developed and includes PD-L1, PD-1, CD8, FoxP3, CD68, a tumor marker
(cytokeratin AE1/3 for non-small cell lung carcinoma, NSCLC), a pan-membrane marker and
DAPI. Through this assay, it is possible to enumerate key cellular subsets and their co-expression
profiles. It is also possible to include spatial parameters, including the distance between PD-1 and
PD-L1, which have not previously been included in predictive or prognostic surgical pathology
specimen-based assays. This mIF assay has increased sensitivity and specificity for response to
anti-PD1 therapy when compared to the assessment of PD-L1 expression alone in multiple tumor
types, including melanoma, NSCLC, and Merkel cell carcinoma, amongst others.
 The purpose of this proposal is to perform inter-site validation of the mIF staining assay
and associated lock-down algorithm amongst four major academic sites (Johns Hopkins, MD
Anderson, Yale University, and Providence Portland Medical Center). Following analytical
validation, discovery and validation cohorts from 250 patients with advanced NSCLC will be used
to establish final assay parameters (including thresholds), linked to clinical outcomes following
anti-PD-1-based therapy. The deliverable of the study is a refined, multiplex biomarker assay for
response/resistance to anti-PD-1 that has been validated across multiple academic sites. The
result will be a multiplex IF assay that is suitably staged for advanced development aimed at
clinical implementation. While NSCLC is the focus of the current grant proposal, preliminary
results suggest that this assay will also have great value in numerous other solid tumor types.

## Key facts

- **NIH application ID:** 10506186
- **Project number:** 1UH2CA272905-01
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Janis M Taube
- **Activity code:** UH2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $224,347
- **Award type:** 1
- **Project period:** 2022-09-16 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10506186

## Citation

> US National Institutes of Health, RePORTER application 10506186, Analytic and Clinical Validation of a 7-plex MIF Assay for Predictive Response of Advanced NSCLC to Anti-PD-1 based Therapy (1UH2CA272905-01). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10506186. Licensed CC0.

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