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1103 Spatial mapping and clinical significance of the CD39/CD73 adenosinergic pathway as a candidate immunotherapy target in non-small cell lung cancer
  1. Kerryan Ashley1,
  2. Krishna Iyer1,
  3. Rakesh Kumar2,
  4. Zachary A Cooper2,
  5. Roy S Herbst1,
  6. Sarah Goldberg1 and
  7. Kurt Schalper1
  1. 1Yale University, New Haven, CT, USA
  2. 2AstraZeneca, Gaithersburg, MD, USA
  • Journal for ImmunoTherapy of Cancer (JITC) preprint. The copyright holder for this preprint are the authors/funders, who have granted JITC permission to display the preprint. All rights reserved. No reuse allowed without permission.


Background The ectonucleotidases CD39 and CD73 act sequentially to convert ATP into highly immunosuppressive adenosine that can accumulate in the extracellular milieu and suppress NK/T-cell anti-tumor responses. Blockade of CD73 in combination with other immunostimulatory antibodies can induce anti-tumor responses in subset of patients with non-small cell lung cancer (NSCLC). The levels, spatial distribution, clinicopathologic associations and biomarker potential of the CD39/CD73 pathway in NSCLC, as well as the impact of chemoradiotherapy, remain poorly understood.

Methods Using multiplexed quantitative immunofluorescence (mQIF), we simultaneously measured the levels of CD39, CD73 protein, CD8+ T-cells and cytokeratin (CK)-expressing cancer-cells in five retrospective NSCLC cohorts represented in tissue microarrays. The mQIF analysis included compartment-based measurements based on fluorescence co-localization as well as single-cell segmentation/phenotyping and spatial analysis. The first three cohorts included baseline tumor samples from patients with stages I-IV NSCLC treated with non-immunotherapy regimens (Cohort #1, n=179; Cohort #2, n=172; Cohort #3, n=267). The fourth cohort (Cohort #4, n= 68) included baseline tumor samples from patients treated with immunotherapy. The fifth cohort (Cohort #5, n=23 pairs) included paired biopsies before and after chemotherapy with or without radiotherapy. Associations between the markers with clinicopathologic variables and outcomes were studied.

Results 90% of NSCLCs showed detectable concurrent expression of both CD39 and CD73, but the levels were highly variable across cases. CD39+ cell density was higher in nonmalignant CK- stromal cell areas and CD73+ cell density was higher within the CK+ cancer cell nests. The levels of the markers were positively associated across the cohorts and high expression of CD39 or CD73 were consistently associated with higher local CD8+ T-cell infiltration and adenocarcinoma histology. CD8+ T-cells were more distant from CD73+ cancer cells than CD73- cancer cells. After chemoradiotherapy, CD73+ cells are significantly depleted. Stromal CD39+ cells and tumor CD73+ cells were associated with better outcomes after immunotherapy.

Conclusions The CD39/CD73 pathway is expressed in the majority of NSCLCs with a distinct tumor/stromal cell distribution. This pathway is associated with local adaptive anti-tumor immune responses, adenocarcinoma histology and better outcome after immunotherapy. Chemo-radiotherapy reduces the density of CD73+ expressing cells in the tumor microenvironment without significantly altering the spatial distribution of these cells relative to CD8+ TILs.

Acknowledgements Funding Source: AstraZeneca

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