Hypoxia is a well-known component of many tumors and acts in an immunosuppressive manner through multiple distinct pathways. 18 F-FMISO PET imaging has been demonstrated to accurately quantify hypoxia in a clinical setting, although it has not been investigated for immunotherapy. Our work demonstrates that effective immunotherapy prevents tumor hypoxia and that uptake of 18 F-FMISO was predictive of subsequent changes in anatomical tumor size. We further demonstrate that PET imaging of hypoxia is correlated with phenotypic characteristics of inflammation through transcriptomics, immune cell spatial correlation, and direct measurement of secreted inflammatory proteins. The upregulation of damage associated molecular pattern signaling in responding tumors warranted investigation of the addition of the hypoxia targeted pro-drug to non-responding tumors in order to enhance checkpoint blockade efficacy. The addition of evofosfamide improved tumor oxygenation and response, providing rationale for immediate clinical investigation of both 18 F-FMISO PET imaging and combination evofosfamide therapy for improved cancer immunotherapy.Research.

The second heart field (SHF) harbors progenitors that are important for heart formation, but little is known about its morphogenesis. We show that SHF population in the mouse splanchnic mesoderm (SpM-SHF) undergoes polarized morphogenesis to preferentially elongate anteroposteriorly. Loss of Wnt5, a putative ligand of the planar cell polarity (PCP) pathway, causes the SpM-SHF to expand isotropically. Temporal tracking reveals that the Wnt5a lineage is a unique subpopulation specified as early as E7.5, and undergoes bi-directional deployment to form specifically the pulmonary trunk and the dorsal mesenchymal protrusion (DMP). In Wnt5a −/− mutants, Wnt5a lineage fails to extend into the arterial and venous poles, leading to both outflow tract and atrial septation defects that can be rescued by an activated form of PCP effector Daam1. We identify oriented actomyosin cables in the medial SpM-SHF as a potential Wnt5a-mediated mechanism that promotes SpM-SHF lengthening and restricts its widening. Finally, the Wnt5a lineage also contributes to the pulmonary mesenchyme, suggesting that Wnt5a/PCP is a molecular circuit recruited by the recently identified cardiopulmonary progenitors to coordinate morphogenesis of the pulmonary airways and the cardiac septations necessary for pulmonary circulation. This article has an associated 'The people behind the papers' interview.

No abstract

<div>AbstractPurpose:<p>Hypoxia is a common characteristic of many tumor microenvironments, and it has been shown to promote suppression of antitumor immunity. Despite strong biological rationale, longitudinal correlation of hypoxia and response to immunotherapy has not been investigated.</p>Experimental Design:<p>In this study, we probed the tumor and its surrounding microenvironment with ¹⁸F-FMISO PET imaging to noninvasively quantify tumor hypoxia <i>in vivo</i> prior to and during PD-1 and CTLA-4 checkpoint blockade in preclinical models of breast and colon cancer.</p>Results:<p>Longitudinal imaging identified hypoxia as an early predictive biomarker of therapeutic response (prior to anatomic changes in tumor volume) with a decreasing standard uptake value (SUV) ratio in tumors that effectively respond to therapy. PET signal correlated with <i>ex vivo</i> markers of tumor immune response including cytokines (IFNγ, GZMB, and TNF), damage-associated molecular pattern receptors (TLR2/4), and immune cell populations (macrophages, dendritic cells, and cytotoxic T cells). Responding tumors were marked by increased inflammation that were spatially distinct from hypoxic regions, providing a mechanistic understanding of the immune signaling pathways activated. To exploit image-guided combination therapy, hypoxia signal from PET imaging was used to guide the addition of a hypoxia targeted treatment to nonresponsive tumors, which ultimately provided therapeutic synergy and rescued response as determined by longitudinal changes in tumor volume.</p>Conclusions:<p>The results generated from this work provide an immediately translatable paradigm for measuring and targeting hypoxia to increase response to immune checkpoint therapy and using hypoxia imaging to guide combinatory therapies.</p></div>

<div>AbstractPurpose:<p>Hypoxia is a common characteristic of many tumor microenvironments, and it has been shown to promote suppression of antitumor immunity. Despite strong biological rationale, longitudinal correlation of hypoxia and response to immunotherapy has not been investigated.</p>Experimental Design:<p>In this study, we probed the tumor and its surrounding microenvironment with ¹⁸F-FMISO PET imaging to noninvasively quantify tumor hypoxia <i>in vivo</i> prior to and during PD-1 and CTLA-4 checkpoint blockade in preclinical models of breast and colon cancer.</p>Results:<p>Longitudinal imaging identified hypoxia as an early predictive biomarker of therapeutic response (prior to anatomic changes in tumor volume) with a decreasing standard uptake value (SUV) ratio in tumors that effectively respond to therapy. PET signal correlated with <i>ex vivo</i> markers of tumor immune response including cytokines (IFNγ, GZMB, and TNF), damage-associated molecular pattern receptors (TLR2/4), and immune cell populations (macrophages, dendritic cells, and cytotoxic T cells). Responding tumors were marked by increased inflammation that were spatially distinct from hypoxic regions, providing a mechanistic understanding of the immune signaling pathways activated. To exploit image-guided combination therapy, hypoxia signal from PET imaging was used to guide the addition of a hypoxia targeted treatment to nonresponsive tumors, which ultimately provided therapeutic synergy and rescued response as determined by longitudinal changes in tumor volume.</p>Conclusions:<p>The results generated from this work provide an immediately translatable paradigm for measuring and targeting hypoxia to increase response to immune checkpoint therapy and using hypoxia imaging to guide combinatory therapies.</p></div>