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Persistent fatigue is a debilitating side effect that impacts a significant proportion of cancer survivors for which there is not yet an FDA-approved treatment. While certainly a multi-factorial problem, persistent fatigue could be due, in part, to associations learned during treatment. Therefore, we sought to investigate the role of associative learning in the persistence of fatigue using a preclinical model of cancer survivorship. For this purpose, we used a murine model of human papilloma virus-related head and neck cancer paired with a curative regimen of cisplatin-based chemoradiation in male C57BL/6J mice. Fatigue-like behavior was assessed by measuring variations in voluntary wheel running using a longitudinal design. Treatment robustly decreased voluntary wheel running, and this effect persisted for more than a month posttreatment. However, when wheels were removed during treatment, to minimize treatment-related fatigue, mice showed a more rapid return to baseline running levels. We confirmed that the delayed recovery observed in mice with continual wheel access was not due to increased treatment-related toxicity, in fact running attenuated cisplatin-induced kidney toxicity. Finally, we demonstrated that re-exposure to a treatment-related olfactory cue acutely re-instated fatigue. These data provide the first demonstration that associative processes can modulate the persistence of cancer-related fatigue-like behavior.

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Progressive tumor growth is associated with deficits in the immunity generated against tumor antigens. Vaccines targeting tumor neoepitopes have the potential to address qualitative defects; however, additional mechanisms of immune failure may underlie tumor progression. In such cases, patients would benefit from additional immune-oncology agents targeting potential mechanisms of immune failure. This study explores the identification of neoepitopes in the MC38 colon carcinoma model by comparison of tumor to normal DNA and tumor RNA sequencing technology, as well as neoepitope delivery by both peptide- and adenovirus-based vaccination strategies. To improve antitumor efficacies, we combined the vaccine with a group of rationally selected immune-oncology agents. We utilized an IL15 superagonist to enhance the development of antigen-specific immunity initiated by the neoepitope vaccine, PD-L1 blockade to reduce tumor immunosuppression, and a tumor-targeted IL12 molecule to facilitate T-cell function within the tumor microenvironment. Analysis of tumor-infiltrating leukocytes demonstrated this multifaceted treatment regimen was required to promote the influx of CD8+ T cells and enhance the expression of transcripts relating to T-cell activation/effector function. Tumor-targeted IL12 resulted in a marked increase in clonality of T-cell repertoire infiltrating the tumor, which when sculpted with the addition of either a peptide or adenoviral neoepitope vaccine promoted efficient tumor clearance. In addition, the neoepitope vaccine induced the spread of immunity to neoepitopes expressed by the tumor but not contained within the vaccine. These results demonstrate the importance of combining neoepitope-targeting vaccines with a multifaceted treatment regimen to generate effective antitumor immunity.

There is no cure for HIV infection, and lifelong antiretroviral therapy (ART) is required. N-803 is an IL-15 superagonist comprised of an N72D mutant IL-15 molecule attached to its alpha receptor and a human IgG1 fragment designed to increase IL-15 activity. Preclinical studies with both HIV and SIV suggest that the drug has potential to reduce virus reservoirs by activating virus from latency and enhancing effector function. We conducted a phase 1 study of N-803 ( NCT02191098 ) in people living with HIV, the primary objective of which was to assess the safety and tolerability of the drug, with an exploratory objective of assessing the impact on peripheral virus reservoirs. ART-suppressed individuals were enrolled into a dose-escalation study of N-803 in four different cohorts (0.3, 1.0, 3.0 and 6.0 mcg kg

This file shows burrowing behavior in tumor-bearing mice that were or were not treated with systemic IL-1ra.

Fatigue is the most common symptom of cancer at diagnosis, yet causes and effective treatments remain elusive. As tumors can be highly inflammatory, it is generally accepted that inflammation mediates cancer-related fatigue. However, evidence to support this assertion is mostly correlational. In this study, we directly tested the hypothesis that fatigue results from propagation of tumor-induced inflammation to the brain and activation of the central proinflammatory cytokine, IL1. The heterotopic syngeneic murine head and neck cancer model (mEER) caused systemic inflammation and increased expression of Il1b in the brain while inducing fatigue-like behaviors characterized by decreased voluntary wheel running and exploratory activity. Expression of Il1b in the brain was not associated with any alterations in motivation, measured by responding in a progressive ratio schedule of food reinforcement, depression-like behaviors, or energy balance. Decreased wheel running occurred prior to Il1b detection in the brain, when systemic inflammation was minimal. Furthermore, mice null for two components of IL1β signaling, the type 1 IL1 receptor or the receptor adapter protein MyD88, were not protected from tumor-induced decreases in wheel running, despite attenuated cytokine action and expression. Behavioral and inflammatory analysis of four additional syngeneic tumor models revealed that tumors can induce fatigue regardless of their systemic or central nervous system inflammatory potential. Together, our results show that brain IL1 signaling is not necessary for tumor-related fatigue, dissociating this type of cancer sequela from systemic cytokine expression.Significance: These findings challenge the current understanding of fatigue in cancer patients, the most common and debilitating sequela associated with malignancy. Cancer Res; 78(3); 695–705. ©2017 AACR.

This file contains methods and references for data presented in the supplement and legends for Supplementary Figures S1-S4

PDF file, 68K, Primary murine tonsil epithelia (1oMTE), MTEs stably expressing HPV16 E6 and E7 together with Ras and luciferase (MEERL), were analyzed by western blot as indicated.

Most squamous cell carcinomas of the head and neck (HNSCC) overexpress ERBB1/EGFR, but EGF receptor (EGFR)-targeted therapies have yielded disappointing clinical results in treatment of this cancer. Here, we describe a novel interaction between EGFR and the ligand EphrinB1 (EFNB1), and we show that EFNB1 phosphorylation and downstream signaling persists in the presence of cetuximab. Mechanistically, cetuximab drives a shift in EGFR dimerization partners within the signaling complex, suggesting that targeted drugs may trigger partner rearrangements that allow persistent pathway activation. EFNB1 attenuation slowed tumor growth and increased survival in a murine model of HNSCC, suggesting a substantial contribution of EFNB1 signaling to HNSCC development. Together, our findings suggest that EFNB1 is part of the EGFR signaling complex and may mediate drug resistance in HNSCC as well as other solid tumors. Cancer Res; 73(18); 5787–97. ©2013 AACR.

Neonatal sepsis is a major cause of morbidity and mortality in neonates and is challenging to diagnose. Infants manifest nonspecific clinical signs in response to sepsis; these signs may be caused by noninfectious conditions. Time to antibiotics affects neonatal sepsis outcome, so clinicians need to identify and treat neonates with sepsis expeditiously. Clinicians use serum biomarkers to measure inflammation and infection and assess the infant's risk of sepsis. However, current biomarkers lack sufficient sensitivity or specificity to be consider useful diagnostic tools. Continued research to identify novel biomarkers as well as novel ways of measuring them is sorely needed.

The oncotherapeutic promise of IL-15, a potent immunostimulant, is limited by a short serum t1/2. The fusion protein N-803 is a chimeric IL-15 superagonist that has a >20-fold longer in vivo t1/2 versus IL-15. This phase 1 study characterized the pharmacokinetic (PK) profile and safety of N-803 after s.c. administration to healthy human volunteers. Volunteers received two doses of N-803, and after each dose, PK and safety were assessed for 9 d. The primary endpoint was the N-803 PK profile, the secondary endpoint was safety, and immune cell levels and immunogenicity were measures of interest. Serum N-803 concentrations peaked 4 h after administration and declined with a t1/2 of ∼20 h. N-803 did not cause treatment-emergent serious adverse events (AEs) or grade ≥3 AEs. Injection site reactions, chills, and pyrexia were the most common AEs. Administration of N-803 was well tolerated and accompanied by proliferation of NK cells and CD8+ T cells and sustained increases in the number of NK cells. Our results suggest that N-803 administration can potentiate antitumor immunity.

Background: Chemoradiotherapy (CRT) for locally-advanced head and neck squamous cell carcinoma (LA-HSNCC) yields 5-year survival rates near 50% despite causing significant toxicity. Dichloroacetate (DCA), a pyruvate dehydrogenase kinase metabolic inhibitor, reduces tumor lactate production and has been used in cancer therapy previously. The safety of adding this agent to CRT is unknown. Methods: Our randomized, placebo-controlled, double-blind phase II study added DCA to cisplatin-based CRT in patients with LA-HNSCC. The primary endpoint was safety by adverse events (AEs). Secondary endpoints compared efficacy via 3-month end-of-treatment response, 5-year progression-free and overall survival. Translational research evaluated pharmacodynamics of serum metabolite response. Results: 45 participants (21 DCA, 24 Placebo) were enrolled from May 2011-April 2014. Higher rates of all-grade drug related fevers (43% vs 8%, p = 0.01) and decreased platelet count (67% vs 33%, p = 0.02) were seen in DCA versus placebo. However, there were no significant differences in grade 3/4 AE rates. Treatment compliance to DCA/placebo, radiation therapy, and cisplatin showed no significant difference between groups. While end-of-treatment complete response rates were significantly higher in the DCA group compared to placebo (71.4% vs 37.5%, p = 0.0362), survival outcomes were not significantly different between groups. Treatment to baseline metabolites demonstrated a significant drop in pyruvate (0.47, p Conclusions: Adding DCA to cisplatin-based CRT appears safe with no detrimental effect on survival and expected metabolite changes compared to placebo. This supports further investigation into combining metabolic agents to CRT.Trial registration number: NCT01386632, Date of Registration: July 1, 2011.

Chemoradiotherapy (CRT) for locally-advanced head and neck squamous cell carcinoma (LA-HSNCC) yields 5-year survival rates near 50% despite causing significant toxicity. Dichloroacetate (DCA), a pyruvate dehydrogenase kinase metabolic inhibitor, reduces tumor lactate production and has been used in cancer therapy previously. The safety of adding this agent to CRT is unknown. Our randomized, placebo-controlled, double-blind phase II study added DCA to cisplatin-based CRT in patients with LA-HNSCC. The primary endpoint was safety by adverse events (AEs). Secondary endpoints compared efficacy via 3-month end-of-treatment response, 5-year progression-free and overall survival. Translational research evaluated pharmacodynamics of serum metabolite response. 45 participants (21 DCA, 24 Placebo) were enrolled from May 2011-April 2014. Higher rates of all-grade drug related fevers (43% vs 8%, p = 0.01) and decreased platelet count (67% vs 33%, p = 0.02) were seen in DCA versus placebo. However, there were no significant differences in grade 3/4 AE rates. Treatment compliance to DCA/placebo, radiation therapy, and cisplatin showed no significant difference between groups. While end-of-treatment complete response rates were significantly higher in the DCA group compared to placebo (71.4% vs 37.5%, p = 0.0362), survival outcomes were not significantly different between groups. Treatment to baseline metabolites demonstrated a significant drop in pyruvate (0.47, p 0.005) and lactate (0.61, p 0.005) in the DCA group. Adding DCA to cisplatin-based CRT appears safe with no detrimental effect on survival and expected metabolite changes compared to placebo. This supports further investigation into combining metabolic agents to CRT. Trial registration number: NCT01386632, Date of Registration: July 1, 2011.

Evasion of the host immune responses is critical for both persistent human papillomavirus (HPV) infection and associated cancer progression. We have previously shown that expression of the homeostatic chemokine CXCL14 is significantly downregulated by the HPV oncoprotein E7 during cancer progression. Restoration of CXCL14 expression in HPV-positive head and neck cancer (HNC) cells dramatically suppresses tumor growth and increases survival through an immune-dependent mechanism in mice. Although CXCL14 recruits natural killer (NK) and T cells to the tumor microenvironment, the mechanism by which CXCL14 mediates tumor suppression through NK and/or T cells remained undefined. Here we report that CD8+ T cells are required for CXCL14-mediated tumor suppression. Using a CD8+ T-cell receptor transgenic model, we show that the CXCL14-mediated antitumor CD8+ T-cell responses require antigen specificity. Interestingly, CXCL14 expression restores major histocompatibility complex class I (MHC-I) expression on HPV-positive HNC cells downregulated by HPV, and knockdown of MHC-I expression in HNC cells results in loss of tumor suppression even with CXCL14 expression. These results suggest that CXCL14 enacts antitumor immunity through restoration of MHC-I expression on tumor cells and promoting antigen-specific CD8+ T-cell responses to suppress HPV-positive HNC.

e13545 Background: The ASCO Quality Oncology Practice Initiative (QOPI) was established in 2006 to facilitate quality measurement and continuous improvement of cancer care.1 ASCO and QOPI leadership encourage a continuous learning environment to develop new tools and methodologies that improve quality and maximize outcomes and value in oncology practice.2 Parallel advances in tumor molecular profiling, bioinformatics, and EMR data curation are adding new insights to support the real-world research of longitudinal patient journeys. Methods: Referencing the QOPI Certification Track 2021 Measures Summary, we employed data visualization techniques to create a precision oncology dashboard for breast cancer. This evaluation included 409 breast cancer patients for whom genomic testing was performed and presented at the Avera molecular tumor board. Patients were then matched to appropriate biomarker-based targeted therapies. Results: The dashboard identified and prioritized areas for enhanced data capture to inform strategies for care-coordination, research, and delivery of specialized care. Conclusions: An interactive, visual platform may help facilitate the integration of QOPI reporting into CancerLinQ, thereby fostering a real-time, bidirectional system for quality reporting, clinical decision support, and rapid learning.3,4 The next stage will (1) expand the tumor list; and (2) apply NLP methods to assess accuracy, outcomes and temporal reasoning to create interactive graphs for patient referral patterns and surgery decision drivers. References: 1. Neuss MN, Desch CE, McNiff KK, et al: A process for measuring the quality of cancer care: The Quality Oncology Practice Initiative. J Clin Oncol. 2005 2. Blayney DW, McNiff K, Eisenberg PD, Gilmore T, Jacobsen PB, Jacobson JO, Kadlubek PJ, Neuss MN, Simone J. Development and future of the American Society of Clinical Oncology's Quality Oncology Practice Initiative. J Clin Oncol. 2014 3. Blayney DW: Enhancing quality through innovation: American Society of Clinical Oncology presidential address 2010. J Clin Oncol. 2010 4. Schilsky R, Hauser R, Mann J, et al: Lessons learned from the development of the CancerLinQ prototype: Clinical decision support. J Clin Oncol. 31, 2013[Table: see text]

TPS6605 Background: Precision medicine represents a new era in oncology and population health. Comprehensive molecular profiling and hereditary cancer testing are increasingly important in the routine management of many malignancies, yet may not be widely adopted in many practices and leads to patients not consistently receiving care recommended by current national guidelines such as NCCN and ASCO. In addition, available targeted DNA sequencing panels may be limited to only defined drivers and lack detection in precancerous conditions. Whole exome sequencing (WES) combined with whole transcriptome sequencing (WTS) represent an expanded testing approach that identifies additional genetic changes beyond standard commercial panel tests. Current evidence suggests that WES/WTS is feasible to carry out in routine clinical practice, with improved detection of somatic alterations compared to smaller panels, and the promise to enhance access to targeted therapies and clinical trials. The primary purpose of the ASAP study is to understand the breadth of molecular characteristics present in participants cared for in a large integrated community-based health system, by linking comprehensive molecular profiles with clinical data extracted and curated from the electronic medical record. Methods: Avera Health is a large, Midwestern, predominantly rural vertically integrated health system with six regional cancer centers and over 40 oncology outreach sites across South Dakota and surrounding states. The ASAP study was initiated in November 2021 in GYN and as of February, the study is enrolling in the Otolaryngology clinic and expected to open in medical oncology and hematology in March/April 2022. The study will expand to include additional cancer centers outside of Sioux Falls later in 2022 and is anticipated to be open for accrual for 5 years. Enrolled participants are expected to receive WES/WTS and proteomics on their primary tumor and metastatic sites were feasible as well as hereditary testing, pharmacogenomics (PGx) and microbiome profiling. Liquid biopsies will be performed every 3 months while on treatment or when treatment is changed, in follow up to coincide with scheduled radiographic testing, and as part of annual routine surveillance. For participants not diagnosed with cancer, each participant will receive hereditary testing, PGx, microbiome, and a liquid biopsy at least annually. Participants also consent to sharing their electronic health information. Future stages of this work will employ natural language processing (NLP) methods to develop algorithms to help identify opportunities to improve patient outcomes. Projected enrollment is 1000 participants in year 1, 1500 participants in year 2, and up to 3000 annually thereafter. Approximately 50 patients have consented as of February 1 2022. Clinical trial information: NCT05142033.

e15036 Background: Advances in tumor molecular profiling and bioinformatics are adding new insights to support real-world research. In this study, paired tumor NGS testing data was curated and analyzed from electronic medical records (EMR) to understand mutational and targeted therapy landscape. Methods: A database initiated in 2013 to collect data, including outcomes, on all patients referred to the molecular tumor board at the Avera Cancer Institute was curated and analyzed. A data pipeline that combined results from curated NGS data with curated EMR data from Avera was used to correlate targetable mutations to a corresponding therapy and assessed treatment patterns. Results: Data were from 1804 patients with 44 cancer types including breast (462; 26%), lung (338;19%), colorectal (198;11%), ovarian (168;9%), and uterine (100;5%). Overall, 83% of patients had a positive NGS result and 67% had a targetable mutation per NCCN guidelines. 32% of patients with a targetable mutation received targeted therapy per guideline. Overall, 381 patients with 28 tumor types received targeted therapies based on NGS results and targetable biomarkers. Of these, 288/381 (76%) were stage III-IV and 93/381 (24%) early stage/stage unknown. In non-small cell lung cancer, 76% of stage III-IV patients with mutations in EGFR (exon 19 deletion, L858, L861, S768, G719), MET (exon 14 skipping, amplification), BRAF V600 and ALK/ ROS1/ RET fusions received a targeted therapy per NCCN guidelines. In advanced breast cancer, 97% with ERBB2 amplification received targeted therapy (trastuzumab, pertuzumab, neratinib, lapatinib, tucatinib). 8% of advanced breast cancer patients with PIK3CA mutation received alpelisib, with all patients receiving the drug since 2019. 38% advanced breast cancer patients with BRCA1/2 mutation received Olaparib. In ovarian cancer, 48% of advanced stage patients received targeted therapy for BRCA 1/2, ATM, CDK12, CHEK2, ERBB2, FANCL, KRAS, ALK, BRAF alterations. Olaparib was used in 70% of advanced prostate cancer patients with BRCA1/2 mutations. Dabrafenib combined with Trametinib were most preferred for BRAF V600 mutation in melanoma and colorectal tumors respectively. Off label use of targeted therapy was observed in about 6% of advanced patients; examples include trametinib for KRAS/NRAS mutation [non-small cell lung cancer, colorectal, pancreatic, ovarian], PARP inhibitors for BRCA1/2, ATM mutations [urothelial, peritoneal], anti-ERBB2 inhibitors for ERBB2 in-frame, missense, exon 20 insertion mutations [lung and breast]. Conclusions: These study results demonstrate that automated data mining can provide insights into the genomic and therapeutic landscape of a real-world patient population. Across all tumor types, most patients are being given appropriate targeted therapies as per guidelines. Off-label use of targeted therapy was also observed.

Failed T cell-based immunotherapies in the presence of genomic alterations in antigen presentations pathways may be overcome by NK cell-based immunotherapy. This approach may still be limited by the presence of immunosuppressive myeloid populations. Here we demonstrate that NK cells (haNKs) engineered to express a PD-L1 chimeric antigen receptor (CAR) haNKs killed a panel of human and murine head and neck cancer cells at low effector-to-target ratios in a PD-L1-dependent fashion. Treatment of syngeneic tumors resulted in CD8 and PD-L1-dependent tumor rejection or growth inhibition and a reduction in myeloid cells endogenously expressing high levels of PD-L1. Treatment of xenograft tumors resulted in PD-L1 dependent tumor growth inhibition. PD-L1 CAR haNKs reduced levels of macrophages and other myeloid cells endogenously expressing high PD-L1 in peripheral blood from patients with head and neck cancer. The clinical study of PD-L1 CAR haNKs is warranted.

PURPOSE Pembrolizumab is a humanized monoclonal antibody that blocks interaction between programmed death receptor-1 (PD-1) and its ligands (PD-L1, PD-L2). Although pembrolizumab is approved for recurrent/metastatic head and neck squamous cell carcinoma (HNSCC), its role in the management of locally advanced (LA) disease is not defined. We report a phase IB study evaluating the safety and efficacy of adding pembrolizumab to cisplatin-based chemoradiotherapy in patients with LA HNSCC. PATIENTS AND METHODS Eligible patients included those with oral cavity (excluding lip), oropharyngeal, hypopharyngeal, or laryngeal stage III to IVB HNSCC (according to American Joint Committee on Cancer, 7th edition, staging system) eligible for cisplatin-based, standard-dose (70 Gy) chemoradiotherapy. Pembrolizumab was administered concurrently with and after chemoradiotherapy with weekly cisplatin. Safety was the primary end point and was determined by incidence of chemoradiotherapy adverse events (AEs) and immune-related AEs (irAEs). Efficacy was defined as complete response (CR) rate on end-of-treatment (EOT) imaging or with pathologic confirmation at 100 days postradiotherapy completion. Key secondary end points included overall (OS) and progression-free survival (PFS). RESULTS The study accrued 59 patients (human papillomavirus [HPV] positive, n = 34; HPV negative, n = 25) from November 2015 to October 2018. Five patients (8.8%) required discontinuation of pembrolizumab because of irAEs, all of which occurred during concurrent chemoradiotherapy; 98.3% of patients completed the full planned treatment dose (70 Gy) of radiotherapy without any delays ≥ 5 days; 88.1% of patients completed the goal cisplatin dose of ≥ 200 mg/m2. EOT CR rates were 85.3% and 78.3% for those with HPV-positive and -negative HNSCC, respectively. CONCLUSION Pembrolizumab in combination with weekly cisplatin-based chemoradiotherapy is safe and does not impair delivery of curative radiotherapy or chemotherapy in HNSCC. Early efficacy data support further investigation of this approach.

Wireless capsule endoscopy (WCE) has revolutionized the capacity for evaluation of the gastrointestinal (GI) tract, but its evaluation is limited to the mucosal surface. To overcome this, ultrasound capsule endoscopy (UCE) that can evaluate the deeper structures beyond the mucosal surface has been proposed and several studies focusing on technology development have demonstrated promising results. However, investigations of the potential for clinical utility of this technology are lacking. This work had two main goals: perform ex vivo and in vivo imaging studies in a swine model to (1) evaluate if acoustic coupling between a capsule with a specific size and GI tract can be achieved only through peristalsis autonomously without any human control and (2) identify key issues and challenges to help guide further research. The images acquired in these studies were able to visualize the wall of the GI tract as well as the structures within demonstrating that achieving adequate acoustic coupling through peristalsis is possible. Critical challenges were identified including level of visualization and area of coverage; these require further in-depth investigation before potential clinical utility of UCE technology can be concluded.

Aortic stenosis provides a unique management challenge during pregnancy and has historically been associated with a high maternal mortality risk. We describe our experience with pregnancies in patients with aortic stenosis at a single center over a 13 year period. When managed by an experienced, multidisciplinary, cardio-obstetrics team, these pregnancies are more likely to reach term with good outcomes.

Human papillomavirus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) is a fast-growing health problem in the United States. By analyzing the transcriptomes of HPV-positive (HPV+) and HPV-negative (HPV-) HNSCC patient tissues, we previously revealed that expression of CXCL14, constitutively expressed in basal epithelial cells, is downregulated in HPV-positive HNSCC. In contrast, the expression of proinflammatory chemokines, particularly CXCL1, CXCL2, and CXCL8, are significantly upregulated in both HPV+ and HPV- HNSCCs. We further showed that restored expression of CXCL14 in HPV+ HNSCC cells leads to tumor-free survival in vivo by increasing MHC-I antigen presentation on the cancer cell surface and CD8+ T cell activation. Interestingly, high CXCL1, CXCL2, and CXCL8 expression levels significantly correlate to poor HNSCC patient survival, showing an exact opposite pattern of the clinical outcomes associated with the CXCL14 expression levels. To determine the roles of these chemokines in HPV+ HNSCC development, CXCL1 and CXCL2 genes (no murine CXCL8 ortholog) were deleted using a CRISPR-Cas9 knockout system in HPV+ murine HNSCC cells. Using these stably engineered HNSCC cells, we investigated tumor growth, immune cell infiltration, and metastasis in immunocompetent syngeneic mice. Interestingly, the deletion of CXCL1 and CXCL2 significantly delays in vivo tumor growth, decreases myeloid-derived suppressor cell (MDSC) infiltration in the tumor, and decreases lung metastasis. However, the numbers of natural killer (NK), CD4+, and CD8+ T cells in the tumor microenvironment were not affected by CXCL1 and CXCL2 knockout. Next, mice injected with the murine HNSCC cells were treated twice daily with AZD5068, a selective inhibitor of CXCR2, which is a common receptor of CXCL1, CXCL2, and CXCL8 expressed on myeloid cells. Our result showed that the treatment of CXCR2 suppresses in vivo tumor growth in a similar pattern with CXCL1 and CXCL2 knockout. These results suggest that the dysregulation of chemokine expression in HPV-infected cells plays an important role in regulating tumor growth, antitumor immune responses, and metastasis of HPV+ HNSCC. Our findings may provide promising strategies to develop novel immunotherapies for HPV+ HNSCC patients which can likely be extended to other non-viral cancers with immunosuppressive phenotypes. Citation Format: Daniel Vermeer, Joseph Westrich, Paul Colbert, Craig Welbon, John H. Lee, David Raben, William C. Spanos, Dohun Pyeon. Chemokine dysregulation creates the immunosuppressive tumor microenvironment and promotes human papillomavirus-associated head and neck cancer [abstract]. In: Abstracts: AACR Virtual Special Conference: Tumor Immunology and Immunotherapy; 2021 Oct 5-6. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(1 Suppl):Abstract nr P042.

Patients with densely innervated tumors suffer with increased metastasis and decreased survival as compared to those with less innervated tumors. We hypothesize that in some tumors, nerves are acquired by a tumor-induced process, called axonogenesis. Here, we use PC12 cells as an in vitro neuronal model, human tumor samples and murine in vivo models to test this hypothesis. When appropriately stimulated, PC12 cells extend processes, called neurites. We show that patient tumors release vesicles, called exosomes, which induce PC12 neurite outgrowth. Using a cancer mouse model, we show that tumors compromised in exosome release are less innervated than controls. Moreover, in vivo pharmacological blockade of exosome release similarly attenuates tumor innervation. We characterize these nerves as sensory in nature and demonstrate that axonogenesis is potentiated by the exosome-packaged axonal guidance molecule, EphrinB1. These findings indicate that tumor released exosomes induce tumor innervation and exosomes containing EphrinB1 potentiate this activity., Tumors can induce axonogenesis which is commonly associated with worse outcomes. Here, the authors show that this phenomenon is mediated by cancer exosomes.

Concentration measurement of particles in suspension is an important procedure performed in biological and clinical laboratories. Existing methods based on instruments such as hemocytometers, Coulter counters and flow cytometers are often laborious, destructive and incapable of in vivo measurements. On the other hand, an ultrasound-based method can be non-destructive and non-invasive and have the potential for in vivo measurement. In this work, a method is presented that estimates absolute particle concentration from high-frequency B-mode ultrasound images of a sample. The method is based on the detection and characterization of the echoes from individual particles to estimate the effective slice thickness of the image. Calibration using a reference sample is not required because the estimation is entirely image based. The particle type differential is also performed by using the backscatter coefficient of each detected echoes. The method is demonstrated by measuring microsphere suspensions as well as human T-cell suspensions. The proposed method has a wide range of potential clinical applications including non-invasive measurement of cell concentration in biological fluids such as cerebrospinal fluid.

HPV-positive head and neck squamous cell carcinoma (HNSCC) is increasingly frequent. Management is particularly debated in the case of postsurgical high-risk features, that is, positive surgical margins and extracapsular spread (ECS). In this increasingly complex emerging framework of HNSCC treatment, representative preclinical models are needed to support future clinical trials and advances in personalized medicine. Here, we present an immunocompetent mouse model based on the implantation of mouse tonsil epithelial HPV16-E6/E7-expressing cancer cells into the submental region of the floor-of-the-mouth. Primary tumors were found to replicate the patterns of human HNSCC local invasion and lymphatic dissemination. To study disease progression after surgery, tumors were removed likely leaving behind residual disease. Surgical resection of tumors was followed by a high rate of local recurrences (>90%) within the first 2-3 weeks. While only 50% of mice had lymph node metastases (LNM) at time of primary tumor excision, all mice with recurrent tumors showed evidence of LNM. To study the consecutive steps of LNM progression and distant metastasis development, LNs from tumor-bearing mice were transplanted into naive recipient mice. Using this approach, transplanted LNs were found to recapitulate all stages and relevant histological features of regional metastasis progression, including ECS and metastatic spread to the lungs. Altogether, we have developed an immunocompetent HPV-positive HNSCC mouse model of postsurgical local recurrence and regional and distant metastasis progression suitable for preclinical studies.

Studies focused on the killing of activated B-RAF melanoma cells by the histone deacetylase (HDAC) inhibitor AR42. Compared to other tumor cell lines, PDX melanoma isolates were significantly more sensitive to AR42-induced killing. AR42 and the multi-kinase inhibitor pazopanib interacted to activate: an eIF2α-Beclin1 pathway causing autophagosome formation; an eIF2α-DR4/DR5/CD95 pathway; and an eIF2α-dependent reduction in the expression of c-FLIP-s, MCL-1 and BCL-XL. AR42 did not alter basal chaperone activity but increased the ability of pazopanib to inhibit HSP90, HSP70 and GRP78. AR42 and pazopanib caused HSP90/HSP70 dissociation from RAF-1 and B-RAF that resulted in reduced 'RAF' expression. The drug combination activated a DNA-damage-ATM-AMPK pathway that was associated with: NFκB activation; reduced mTOR S2448 and ULK-1 S757 phosphorylation; and increased ULK-1 S317 and ATG13 S318 phosphorylation. Knock down of PERK, eIF2α, Beclin1, ATG5 or AMPKα, or expression of IκB S32A S36A, ca-mTOR or TRX, reduced cell killing. AR42, via lysosomal degradation, reduced the protein expression of HDACs 2/5/6/10/11. In vivo, a 3-day exposure of dabrafenib/trametinib resistant melanoma cells to the AR42 pazopanib combination reduced tumor growth and enhanced survival from ~25 to ~40 days. Tumor cells that had adapted through therapy exhibited elevated HGF expression and the c-MET inhibitor crizotinib enhanced AR42 pazopanib lethality in this evolved drug-resistant population.

Background: Mammary type-myofibroblastoma is a rare, benign tumor that is morphologically identical to myofibroblastoma of the breast. We describe, for the first time, a case of mammary-type myofibroblastoma arising in the larynx. Methods and results: A 70 year-old man presented with dysphagia and dysphonia and was found to have a right vocal cord cystic-appearing lesion, which was surgically excised. Histologic examination showed short fascicles of spindle cells with eosinophilic cytoplasm, oval nuclei with fine chromatin and inconspicuous nucleoli, thick bundles of collagen, and peripherally located focal scattered adipocytes. Immunohistochemical staining was positive for CD34 and negative for MDM2, CDK4, ER, and PR. Conclusion: We report the first rare case of a patient with a mammary-type myofibroblastoma of the larynx. While rare, it is important to consider this in the differential diagnosis when a spindle-cell tumor arises in the head and neck and to review the histopathological findings. Keywords: Myofibroblastoma, Vocal cord tumor, Microdirect laryngoscopy, Histology, Larynx

In the USA, maternal mortality has been rising since the 1980s. Cardiovascular disease is recognized as the leading cause of this worrisome trend, and a multidisciplinary approach to the care of patients with cardiovascular conditions during pregnancy is becoming increasingly important. We outline the literature supporting this multidisciplinary approach, highlight our center’s experience in building and expanding an integrated cardio-obstetrics practice, and provide guidance regarding patient selection and management within a combined practice. Antenatal management patterns and delivery planning for patients with cardiovascular disease during pregnancy vary substantially among cardiovascular and obstetric and maternal fetal medicine practices in the USA. The need for multidisciplinary care between cardiologists and obstetricians is evident and has been supported by best practice statements from the American Heart Association, the American College of Obstetrics and Gynecology, and the Cardiac Disease in Pregnancy Study (CARPREG) investigators, whose CARPREG II risk score included “late first antenatal visit” as a predictor of adverse outcomes of pregnancy. We have solid evidence supporting a multidisciplinary approach to the care of patients with cardiac conditions in pregnancy. This approach is optimal because it facilitates a consistent and clear message to the patient (and those caring for each patient) regarding management and risks associated with pregnancy, as well as subsequent risk and postpartum follow-up. We support the extension of clinical collaboration between obstetricians and cardiologists to the research realm and know that working together to investigate the outcomes of moms with heart conditions and their babies will provide clinically meaningful information to support the care of these unique patients.

Meningiomas are the most common adult primary tumor of the central nervous system, but there are no known effective medical therapies for recurrent meningioma, particularly for World Health Organization grade II and III tumors. Meningiomas arise from the meninges, located outside the blood-brain barrier, and therefore may be directly targeted by antibody-mediated immunotherapy. We found that programmed cell death ligand 1 (PD-L1) was highly expressed in multiple human malignant meningioma cell lines and patient tumor samples. PD-L1 was targeted with the anti–PD-L1 antibody avelumab and directed natural killer cells to mediate antibody-dependent cellular cytotoxicity (ADCC) of PD-L1–expressing meningioma tumors both in vitro and in vivo. ADCC of meningioma cells was significantly increased in target cells that upregulated PD-L1 expression and, conversely, abrogated in tumor cells that were depleted of PD-L1. Additionally, the high-affinity natural killer cell line, haNK, outperformed healthy donor NK cells in meningioma ADCC. Together, these data support a clinical trial designed to target PD-L1 with avelumab and haNK cells, potentially offering a novel immunotherapeutic approach for patients with malignant meningioma.