Faculty Bibliography

PURPOSE/UNASSIGNED:Recurrence in early-stage laryngeal carcinoma after radiation therapy often necessitates a total salvage laryngectomy (TSL). Understanding the genetic landscape that influences cancer progression after radiation therapy may help future treatments. We investigated early-stage laryngeal carcinoma patients treated with radiotherapy to identify mutation patterns that predispose to disease recurrence. METHODS/UNASSIGNED:The patient cohort had 35 early-stage laryngeal head and neck squamous cell carcinoma (HNSCC) patients (T1 = 15, T2 = 20) treated with radiotherapy (RT). We stratified patients: Responders (no recurrence, N = 14), non-responders (recurrence within 12 months, N = 21), and non-responders undergoing TSL (N = 18). We employed whole exome sequencing to characterize gene mutations using the Genome Analysis Toolkit (GATK) for variant detection and impact on critical biological pathways and post-radiotherapy patient outcomes. Pathway analysis Ingenuity and Reactome Pathway Analysis tools were used to explore the mutated gene pathways. RESULTS –/UNASSIGNED:Differential mutation analysis was performed in the respective groups to find driver genes. In the pre-treatment samples, KCNT2 and AGAP6 mutations were exclusively found in non-responders (OR = 0, P = 0.005 and OR = 0, P = 0.027, respectively), while ADAMTS7 mutations were solely present in responders (OR = inf, P = 0.019). PLEC mutations were more prevalent in responders (OR = 11.6, P = 0.006). Pathway analysis revealed that significant genes were involved in the RND2 GTPase cycle, protein O-glycosylation-related diseases, and apoptotic pathways. Post-treatment analysis in patients undergoing TSL had enrichment of mutations in apoptosis regulation pathways. CONCLUSIONS –/UNASSIGNED:The study reveals that mutations in apoptosis controlling genes were predominantly represented in the larynx non-responder patients both in the pre-radiotherapy and post-TSL populations.

OBJECTIVE:To define oncologic outcomes in Veterans in the modern era using a multi-institutional cohort designed to support development and validation of prognostic and predictive biomarkers for oropharyngeal squamous cell carcinoma (OPSCC). METHODS:A retrospective analysis was conducted including adult OPSCC patients treated at one of nine Veterans Affairs Medical Centers between 2000 and 2024; inclusive of 597 HPV-associated and 197 HPV-independent tumors. All patients were treated with curative intent external beam radiotherapy (100%) with (90%) or without concurrent chemotherapy. RESULTS:A total of 894 adult patients (mean age, 64 years; 881 (99.5%) male) were included in the study; 22% of patients self-identified as Black. The estimated 2- and 5-year OS rates for the entire cohort were 71% and 54%, respectively and lagged substantially behind locoregional control (LRC) and distant metastatic control (DMC). For Veterans with HPV-associated OPSCC, LRC and DMC at 5 years were 87% and 87% respectively. The strongest drivers of OS and LRC were T-classification and chemotherapy choice on univariate and multivariate analysis. CONCLUSIONS:Although LRC and DMC rates among Veterans track well with recently completed clinical trial outcomes, OS rates lag substantially suggestive of higher rates of non-cancer-specific mortality. Together, these data suggest that predictive biomarker strategies focused on treatment effectiveness should be predicated on LRC and DMC rather than OS. This multicenter study is the first step in providing a robust dataset capable of developing and optimizing artificial intelligence (AI)-informed prognostic and predictive strategies essential to a precision oncology approach to OPSCC.

PURPOSE/OBJECTIVE:This prospective, non-randomized phase II single-arm pilot trial aimed to explore favorable mid-treatment nodal response (FMNR) through CT imaging to guide de-escalated chemoradiation therapy (CRT) in patients with favorable risk, node-positive HPV-associated oropharyngeal cancer (OPC). MATERIALS AND METHODS/METHODS:. At week 4, CT imaging evaluated nodal response: ≥40% reduction warranted de-escalation to 60 Gy, while <40% reduction continued standard CRT. Primary endpoint was 2-year PFS from initiation of dose de-escalated CRT. Tissue tumor modified viral (TTMV) HPV DNA samples and DW-MRI were collected at baseline and week 4. MDADI questionnaires were collected at baseline, 1, 3, 6, 12, and 24 months. RESULTS:Of 39 patients, 26 had FMNR and underwent de-escalated treatment. 13 pts had slow mid-treatment nodal shrinkage and received standard dose. At a median follow-up of 47.4 months, the 2-year PFS was 92.1% (95% CI: 0.72-0.98) for the deescalated dose group and 92.3% for the standard dose patients (95% CI: 0.57-0.99), p=0.96. With a median survival follow up of 48.9 months (range: 16.7-77.8 months), there were no deaths or distant failures. FMNR was associated with rapid TTMV HPV DNA clearance, reduced TTMV HPV DNA flare, lower baseline and week 4 MRI diffusivity, and higher baseline and week 4 MRI diffusional kurtosis. No differences in acute or late maximum grade 3-4 toxicity by patient were noted. MDADI composite scores showed minimal clinical important difference (MCID) in the de-escalated group at 1-month post-treatment while the standard group had MCID up to 1-year post-treatment. No patients required feeding tube placement. CONCLUSIONS:De-escalated CRT using CT-based mid-treatment nodal response in favorable risk, node-positive HPV-associated OPC achieved excellent 2-year PFS and OS rates and represents a potential approach in better selecting patients for treatment de-escalation.

Remote access approaches to the neck have gained attention due to cosmetic concerns with conventional cervical incisions. Their safety, reproducibility, and oncologic outcomes remain to be fully validated. We retrospectively analyzed our initial experience with retroauricular robotic neck surgery using the Da Vinci system. Thirty-two patients were included. Data collected comprised demographics, procedure type, surgical features, and oncological follow-up. Thirty-five surgeries were performed: 13 posterolateral neck dissections (levels II-V) with central (VI), 5 modified radical dissections (I-V), 5 posterolateral dissections (II-V), and 1 super-selective neck dissection (I). Twelve neck dissections were combined with thyroidectomy. Additional procedures included 4 partial thyroidectomies, 3 submandibular gland resections, 3 schwannoma resections, and 1 branchial cyst excision, totaling 60 procedures. Twenty patients had malignant disease (62.5%). Median hospital stay was 2 days (range 1 9), similar to conventional approaches. Complications included 5 temporary nerve palsies, 1 lymphatic fistula, 1 transient hypoparathyroidism, and 1 minor flap necrosis, all managed conservatively; 1 hematoma required reoperation. Conversion to a conventional approach occured in 4 cases (6.7%). No additional intraoperative or postoperative complications were observed. The mean lymph node yield was 54 in posterolateral/radical dissections and 13 in central dissections. Median follow-up of malignant cases was 41.3 months (range 18-56), with a 3-year regional control rate of 94.7%. Our experience with 60 retroauricular robotic procedures demonstrates this approach to be safe, feasible, and oncologically effective, with complication rates, hospital stay, and oncological outcomes comparable to conventional surgery.

Epigenetic changes drive gene expression alterations, contributing to oncogenesis and drug resistance. Lysosomes play a key role in cell signaling and sequestering toxins, including chemotherapeutic agents, which are then expelled through lysosomal exocytosis-a process linked to drug resistance. However, the epigenetic regulation of lysosomal exocytosis is poorly understood. We hypothesize that epigenetic modifier drugs (epidrugs) inhibiting this exocytosis could serve as potential cancer therapeutics. To explore this, we screened more than 150 epidrugs targeting various epigenetic proteins for their combined cytotoxic effects with cisplatin, their impact on lysosomal exocytosis, and lysosomal biogenesis. Two type I PRMT inhibitors, MS023 and GSK3368715, showed synergy with cisplatin, reduced cell viability, and inhibited lysosomal exocytosis without altering lysosomal biogenesis gene expression. RNA-seq analysis revealed differentially expressed genes involved in vesicular trafficking and lysosome dynamics, suggesting novel regulatory mechanisms. These inhibitors also synergized with other lysosome-sequestered drugs, indicating a broader application in overcoming drug resistance. Analysis of patient data further linked lower type I PRMT levels to better responses, highlighting their potential as combination therapy candidates to enhance chemotherapy efficacy and improve cancer survival rates.

We leverage a clinical trial (NCT04080804) that compared neoadjuvant anti-PD-1, anti-PD-1+CTLA-4, and anti-PD-1+LAG-3 therapies in head and neck squamous cell carcinoma patients. Combination therapies promote higher pathologic response rates versus monotherapy, and major pathologic response is associated with better survival. To address whether successful immune checkpoint inhibitor (ICI) regimens act through similar or distinct pathways, we robustly and longitudinally characterize transcriptional and proteomic dynamics of CD8⁺ tumor-infiltrating lymphocytes (TILs) in a clonal manner. Anti-PD-1+LAG-3 reprograms CD8⁺ TIL with type-I interferon response and exhaustion gene programs into effector memory and resident memory (T_EM/T_RM). In contrast, anti-PD-1+CTLA-4 activates and expands pre-existing T_EM/T_RM CD8⁺ TIL, but does not rejuvenate exhausted phenotypes into T effector cells. Anti-PD-1+LAG-3, but not anti-PD-1+CTLA-4, induces widespread TCR sharing among the different transcriptional states, as well as increased TCR diversity in responding patients. Our data suggest doublet regimen-specific transcriptional and clonal dynamics of tumor-reactive CD8⁺ T cells.

BACKGROUND:Human papillomavirus (HPV) negative oropharyngeal squamous cell carcinoma (OPSCC) is associated with worse survival when compared to HPV-positive OPSCC. Primary surgery is one option to intensify therapy in this high-risk group of patients. Unfortunately, the only randomized trial to explore this approach (RTOG 1221) failed to accrue and the role of primary surgery in the treatment of HPV-negative OPSCC remains unanswered. METHODS:A systematic review and meta-analysis were performed to examine the outcomes of surgery in the treatment of HPV-negative OPSCC. We used the PRISMA statement for reporting and queried Pubmed, Web of Science and the Cochrane databases for studies examining the use of primary surgery in the treatment of HPV-negative OPSCC. Excluded from analysis were reviews, commentaries, case series with fewer than 10 patients, and studies that included HPV-negative head and neck cancers of mixed sites. Our primary outcomes were 2-year and 5-year overall survival (OS) and disease-free survival (DFS). OS and DFS were pooled using meta-analysis of proportions. RESULTS: = undetermined; 2 studies). CONCLUSIONS:The two- and five-year OS for patients with HPV-negative OPSCC treated with any surgical approach and pathology-directed adjuvant therapy is 84% and 72%, respectively. The two- and five-year OS for HPV-negative OPCSCC treated with transoral surgery and pathology-directed adjuvant therapy is 87% and 82%, respectively.

Communication between intracellular organelles including lysosomes and mitochondria has recently been shown to regulate cellular proliferation and fitness. The way lysosomes and mitochondria communicate with each other (lysosomal/mitochondrial interaction, LMI) is, emerging as a major determinant of tumor proliferation and growth. About 30% of squamous carcinomas (including squamous cell carcinoma of the head and neck, SCCHN) overexpress TMEM16A, a calcium-activated chloride channel, which promotes cellular growth and negatively correlates with patient survival. We have recently shown that TMEM16A drives lysosomal biogenesis, but its impact on mitochondrial function has not been explored. Here, we show that in the context of high TMEM16A SCCHN, (1) patients display increased mitochondrial content, specifically complex I; (2) In vitro and in vivo models uniquely depend on mitochondrial complex I activity for growth and survival; (3) NRF2 signaling is a critical linchpin that drives mitochondrial function, and (4) mitochondrial complex I and lysosomal function are codependent for proliferation. Taken together, our data demonstrate that coordinated lysosomal and mitochondrial activity and biogenesis via LMI drive tumor proliferation and facilitates a functional interaction between lysosomal and mitochondrial networks. Therefore, inhibition of LMI instauration may serve as a therapeutic strategy for patients with SCCHN. Implications: Intervention of lysosome-mitochondria interaction may serve as a therapeutic approach for patients with high TMEM16A expressing SCCHN.