Phase II Window Study of Olaparib Alone or with Cisplatin or Durvalumab in Operable Head and Neck Cancer.

Purpose: We conducted a phase II randomized noncomparative window of opportunity (WOO) trial to evaluate the inhibition of cellular proliferation and the modulation of immune microenvironment after treatment with olaparib alone or in combination with cisplatin or durvalumab in patients with operable head and neck squamous cell carcinoma (HNSCC). Experimental Design: Forty-one patients with HNSCC were randomized to cisplatin plus olaparib (arm A), olaparib alone (arm B), no treatment (arm C) or durvalumab plus olaparib (arm D). The primary endpoint was to evaluate the percentage of patients in each arm that achieved a reduction of at least 25% in Ki67. Secondary endpoints included objective response rate (ORR), safety, and pathologic complete response (pCR) rate. Paired baseline and resection tumor biopsies and blood samples were evaluated for prespecified biomarkers. Results: A decrease in Ki67 of at least 25% was observed in 44.8% of treated patients, as measured by quantitative immunofluorescence. The ORR among treated patients was 12.1%. pCR was observed in 2 patients. Two serious adverse events occurred in 2 patients.Programmed death ligand 1 (PD-L1) levels [combined positive score (CPS)] were significantly higher after treatment in arms A and D. Expression of CD163 and colony-stimulating factor 1 receptor (CSF1R) genes, markers of M2 macrophages, increased significantly posttreatment whereas the expression of CD80, a marker of M1 macrophages, decreased. Conclusion: Preoperative olaparib with cisplatin or alone or with durvalumab was safe in the preoperative setting and led to decrease in Ki67 of at least 25% in 44.8% of treated patients. Olaparib-based treatment modulates the tumor microenvironment leading to upregulation of PD-L1 and induction of protumor features of macrophages. Significance: HNSCC is characterized by defective DNA repair pathways and immunosuppressive tumor microenvironment. PARP inhibitors, which promote DNA damage and "reset" the inflammatory tumor microenvironment, can establish an effective antitumor response. This phase II WOO trial in HNSCC demonstrated the immunomodulatory effects of PARP inhibitor-induced DNA damage. In this chemo-naïve population, PARP inhibitor-based treatment, reduced tumor cell proliferation and modulated tumor microenvironment. After olaparib upregulation of PD-L1 and macrophages, suggests that combinatorial treatment might be beneficial. Synopsis: Our WOO study demonstrates that preoperative olaparib results in a reduction in Ki67, upregulation of PD-L1 CPS, and induction of protumor features of macrophages in HNSCC.

Patterns of Response to Immune Checkpoint Inhibitors in Association with Genomic and Clinical Features in Patients with Head and Neck Squamous Cell Carcinoma (HNSCC).

Background: We sought to compare patterns of response to immune checkpoint inhibitors (ICI) with respect to clinical and genomic features in a retrospective cohort of patients with recurrent/metastatic (R/M) head and neck squamous cell carcinoma (HNSCC). Methods: One hundred seventeen patients with R/M HNSCC treated with ICI were included in this study. Tumor growth kinetics (TGK) prior to and TGK upon immunotherapy (IO) was available for 49 patients. The TGK ratio (TGKR, the ratio of tumor growth velocity before and upon treatment) was calculated. Hyperprogression (HPD) was defined as TGKR ≥ 2. Results: HPD was documented in 18 patients (15.4% of the whole cohort). Patients with HPD had statistically significant shorter progression free survival (PFS) (median PFS 1.8 months (95% CI, 1.03-2.69) vs. 6.1 months for patients with non-HPD (95% CI, 4.78-7.47), p = 0.0001) and overall survival (OS) (median OS 6.53 months (95% CI, 0-13.39) vs. 15 months in patients with non HPD (95% CI, 7.1-22.8), p = 0.0018). In a multivariate Cox analysis, the presence of HPD remained an independent prognostic factor (p = 0.049). Primary site in the oral cavity and administration of ICI in the second/third setting were significant predictors of HPD in multivariate analysis (p = 0.028 and p = 0.012, respectively). Genomic profiling revealed that gene amplification was more common in HPD patients. EGFR gene amplification was only observed in HPD patients, but the number of events was inadequate for the analysis to reach statistical significance. The previously described MDM2 amplification was not identified. Conclusions: HPD was observed in 15.4 % of patients with R/M HNSCC treated with IO and was associated with worse PFS and OS. EGFR amplification was identified in patients with HPD.

Prognostic impact of indoleamine 2,3-dioxygenase 1 (IDO1) mRNA expression on circulating tumour cells of patients with head and neck squamous cell carcinoma.

BACKGROUND: We sought to determine the prognostic role of indoleamine 2,3-dioxygenase 1 (IDO1) by evaluating IDO1 expression in circulating tumour cells (CTCs) at baseline and after completion of chemoradiotherapy in patients with locally advanced (LA) head and neck squamous cell carcinoma (HNSCC) treated with curative intent. METHODS: In a prospective cohort of 113 patients with LA HNSCC, we evaluated expression of IDO1 in the EpCAM+ CTC fraction at baseline and after cisplatin chemoradiation. The prognostic value of combined programmed cell death ligand-1 (PDL-1) and IDO1 expression was assessed. RESULTS: IDO1 was significantly overexpressed at baseline compared with the post-treatment counterparts (p=0.007). IDO1 messenger RNA (mRNA) expression at baseline was associated with better survival in terms of progression-free survival (PFS) (HR=0.19, p=0.017). Post-treatment IDO1 mRNA levels were correlated with unfavourable prognosis in terms of overall survival (OS) (HR=3.27, p=0.008). Patients with combined decreased expression levels of PDL-1 and IDO1 after treatment exhibited superior PFS (p=0.043) and OS (p=0.021). CONCLUSIONS: Our results strongly suggest that IDO1 mRNA expression is an independent prognostic factor for clinical outcome. Our study provides useful information for future trials combining chemoradiation with immune checkpoint inhibitors and IDO1 inhibitors.

Advanced cutaneous squamous cell carcinoma: how is it defined and what new therapeutic approaches are available?

PURPOSE OF REVIEW: Despite the overall excellent survival rates in patients with cutaneous squamous cell carcinoma (cSCC), advanced cutaneous SCCs are associated with high patient morbidity and mortality. Therefore, important unmet clinical needs persist: identifying high risk patients and choosing optimal treatment approaches. RECENT FINDINGS: In recent years, a better understanding of the biology of cSCC and its clinical progression have led to improved staging systems and new promising treatments for advanced disease. Such treatments include PD1 inhibitors, such as cemiplimab, which was recently approved for the treatment of cutaneous SCC, and pembrolizumab whose efficacy in the treatment cSCC is still being investigated. Other treatments, such as epidermal growth factor receptor inhibitors have also been used in the treatment of cSCC with moderate success. Several clinical and histological risk factors are considered key in estimating the risk or recurrence or metastasis in cSCCs and, therefore, influence the appropriate treatment choice and patient monitoring. SUMMARY: The present study reviews the current definition of advanced cSCC and discusses the new systemic approaches, including checkpoint inhibitors.

Surrogates of immunologic cell death (ICD) and chemoradiotherapy outcomes in head and neck squamous cell carcinoma (HNSCC).

OBJECTIVES: Chemoradiation can induce immunogenic (ICD) or tolerogenic cell death. ICD relies on the generation of damage-associated molecular patterns which can stimulate toll-like receptors (TLRs). We sought to determine whether we can predict responses to chemoradiation by measuring surrogate biomarkers of ICD in a cohort of patients with locally advanced (LA) head and neck squamous cell carcinoma (HNSCC). MATERIALS AND METHODS: In a cohort of 113 LA HNSCC pts we evaluated expression of TLR4, TLR7 and TLR9 in the EpCAM + circulating tumor cell (CTC) fraction at baseline and after cisplatin chemoradiation. We also quantified changes in chemokines CXCL10, CXCL16 and IL-2R in the serum. RESULTS: Seventy three patients had evaluable specimens. Among cases with biomarker assessment at baseline and post treatment, 36.8% had an increase in CXCL10 levels (p = 0.022), 73.7% had an increase in CXCL16 levels (p = 0.002) and 63.8% had an increase in IL2Ra levels (p = 0.032) with treatment. 52.0% of evaluable cases at baseline and post-treatment had an increase in TLR4 levels (p = 0.996), 42.9% had an increase in TLR7 levels (p = 0.042) and 27.7% had increase in TLR9 levels (p = 0.011) with treatment. CXCL10 levels at baseline were significantly associated with PFS and OS (p = 0.010 and p = 0.032, respectively). CONCLUSIONS: Our results suggest that chemoradiation leads to quantifiable effects in surrogate markers of ICD. These effects may inform trials combining chemoradiation with immune checkpoint inhibitors. In addition, CXCL10 has prognostic effect in pts treated with chemoradiation.

Papillary cystadenoma of the lower lip exhibiting ciliated pseudostratified columnar epithelium: report of a bizarre case and review of the literature.

BACKGROUND: Salivary gland tumors are uncommon and constitute 2-6.5 % of all head and neck neoplasms. Tumors of minor salivary gland origin account for less than 25 % of all salivary gland neoplasms. Papillary cystadenoma of salivary glands is a rare benign epithelial neoplasm characterized by multicystic growth in which the epithelium exhibits adenomatous proliferation. Papillary cystadenoma of minor salivary glands most frequently involves the lip, buccal mucosa, and palate. This tumor typically presents as a slow-growing, painless mass, usually with diameter of less than 1 cm and clinical resemblance to a mucocele. Although most papillary cystadenomas are predominantly of one cell type, a regional variability may be present. CASE REPORT: We present a case of papillary cystadenoma of the minor salivary glands in a 58-year-old patient exhibiting an upper respiratory tract epithelium, a profoundly atypical benign tumor. DISCUSSION: This type of minor salivary gland tumor epithelium in the lower lip may be the result of a metaplastic process or simply another neoplastic manifestation of papillary cystadenoma. As far as the differential diagnosis of this entity is concerned, it is important to distinguish it from papillary cystadenoma lymphomatosum (Warthin's tumor), low-grade mucoepidermoid carcinoma, the papillary-cystic variant of acinic cell carcinoma, and cystadenocarcinoma Recognition of this lesion is important for the clinician since the differential diagnosis includes lesions with similar clinical appearance and infiltrative behavior.

Traumatic ulcerative granuloma with stromal eosinophilia: a lesion with alarming histopathologic presentation and benign clinical course.

Traumatic ulcerative granuloma with stromal eosinophilia (TUGSE) is a chronic, benign, self-limiting lesion of the oral mucosa. Clinically, the ulceration is characterized by the presence of indurated elevated borders and may resemble pyogenic granuloma or even squamous cell carcinoma of the mouth. Pathogenesis of the lesion is unclear. Although it had been suggested that TUGSE may represent a CD30+ lymphoproliferative disorder, this theory is currently not supported by evidence. We are presenting a classic example of TUGSE, its clinical course, differential diagnosis, and treatment.

Desquamative gingivitis as a manifestation of chronic mucocutaneous disease.

Desquamative gingivitis (DG) is a clinical descriptive term indicating "peeling gums". DG is usually the result of a disease process that causes separation of the epithelium from the underlying connective tissue in the oral masticatory mucosa. DG may be a manifestation of several mucocutaneous diseases, most commonly cicatricial pemphigoid, pemphigus vulgaris and lichen planus. Correct diagnosis of the underlying disease in DG patients requires careful clinical observation, detailed examination of medical history, biopsy and histopathological examination of the lesions as well as more specialized tests such as direct and indirect immunofluorescence. Treatment of DG consists of treating the underlying disease and often requires the use of immunosuppressive agents, such as corticosteroids. Elimination of local gingival irritants, such as dental plaque and calculus, can significantly improve the treatment outcome.

Severe periodontitis in a patient with hyperoxaluria and oxalosis: a case report and review of the literature.

BACKGROUND: Hyperoxaluria is a metabolic disease with excessive urinary oxalate excretion that can be primary or secondary. Hyperoxaluria can result in chronic renal disease and renal failure. Calcium oxalate crystals can be deposited in oral tissues, and the disease can be associated with severe periodontitis and tooth loss. METHODS: The periodontal condition of a 38-year-old patient with a diagnosis of hyperoxaluria and end-stage renal disease is presented. The patient's periodontal status was monitored over a period of several weeks, and extracted teeth were submitted for histopathologic evaluation. RESULTS: The patient was diagnosed with generalized severe periodontitis and external root resorption. Initial periodontal treatment consisting of oral-hygiene instructions and scaling and root planing was performed. However, despite an initial decrease of soft tissue inflammation, the patient's periodontal condition deteriorated, and eventually, all teeth had to be extracted. The deposition of calcium oxalate crystals in the periodontal tissues was confirmed histologically. CONCLUSIONS: Long-standing hyperoxaluria can be associated with severe periodontitis and external root resorption resulting in tooth loss. The pathogenetic mechanisms of hard tissue destruction are still unclear.

Expression of MMP-13 (collagenase-3) in long-term cultures of human dental pulp cells.

UNLABELLED: Matrix metalloproteinase-13 (MMP-13 or collagenase-3) is a member of the family of matrix metalloproteinases (MMPs) produced in high amounts by cells with mineralising potential. Human dental pulp has been shown to express high levels of MMP-13 RNA. OBJECTIVE: Since human dental pulp derived cells (HDPC) are known to possess osteoprogenitor properties, we investigated the pattern of expression of MMP-13 in long-term cultures of those cells under conditions that support mineralisation in vitro. DESIGN: Impacted teeth or teeth extracted for orthodontic purposes were used to obtain dental pulp explants and HDPC were cultured for approximately 5 weeks. Pro- and active MMP-13 levels were determined in the cell culture supernatants by means of enzyme-linked immunosorbent assay (ELISA). Cell growth was evaluated through DNA content and osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity and Alizarin Red staining. RESULTS: Mineralising cultures of HDPC produced significantly higher levels of pro-MMP-13 compared to control cultures. Both pro- and active MMP-13 levels displayed a characteristic peak that was found to coincide with the peak in alkaline phosphatase activity and the onset of mineralisation. Once mineralisation was firmly established, MMP-13 expression was significantly reduced. CONCLUSIONS: Evidence from this study suggests a role for MMP-13 in the transition of human dental pulp cells to a mature mineralising phenotype and points to MMP-13 as a possible marker in HDPC differentiation.

Use of guided tissue regeneration in the treatment of a lateral periodontal cyst with a 7-month reentry.

BACKGROUND: The lateral periodontal cyst (LPC) is an unusual cyst of odontogenic origin, most frequently encountered in the mandible between the roots of canines and premolars. The most common treatment for LPC is surgical enucleation. This article reports a case of an LPC treated with guided tissue regeneration (GTR) and bone allograft. METHODS: A 74-year-old woman presented for periodontal examination. Radiographs revealed a cystic lesion with LPC characteristics. After surgical incisions and flap reflection, the cyst was removed and sent for biopsy. Because of the anatomy of the resulting intrabony defect, GTR was considered the ideal treatment. RESULTS: The biopsy revealed the histologic features of an LPC. Radiographs at 7 months post-treatment indicated bone fill of the initial defect. Although some attachment loss occurred, the reentry demonstrated a high percentage of bone fill of the defect after 7 months. CONCLUSION: Depending on the anatomy of the defect left after the removal of an LPC, GTR, along with bone grafting, can be a very useful tool for its treatment by reducing the attachment loss observed after simple enucleation of the cyst.

Osteogenic differentiation of human mesenchymal bone marrow cells in silk scaffolds is regulated by nitric oxide.

Bone marrow-derived mesenchymal stem cells (BMSC) are a powerful tool for tissue engineering and can be used in the regeneration of bone and other tissues. Nitric oxide (NO) produced by the endothelial NO synthase (eNOS) plays an important role in bone development and healing. We hypothesized that NO plays a role in osteogenic differentiation of BMSC cultured in three-dimensional silk scaffolds. eNOS protein was measured by Western Analysis and its activity was assessed by measuring nitrite in culture supernatants. Mineralization was evaluated through calcium deposition and the expression of genes associated with osteogenic differentiation (collagen I, RUNX2, and osteocalcin) was quantified using real-time RT-PCR. eNOS was consistently expressed with minor fluctuations, but NO production significantly increased at later time points (weeks 4 and 5). Addition of a competitive NOS inhibitor (L-NAME) resulted in a modest decrease in calcium deposition, which became statistically significant in week 5. This was preceded by a dramatic decrease in RUNX2 and osteocalcin expression in week 4. These results support our hypothesis and implicate NO as an important player in bone tissue engineering.

Inhibition of tooth movement by osteoprotegerin vs. pamidronate under conditions of constant orthodontic force.

The undesired movement of anchor teeth, and relapse of previously moved teeth, are major clinical problems in orthodontics. Dental implants are increasingly used to preserve anchorage, but these are costly and require invasive surgical procedures. An alternative strategy for maintaining anchorage may be the use of biological inhibitors of osteoclastic bone resorption. In the present study, we investigated the relative efficacy of pamidronate vs. osteoprotegerin (OPG) in inhibiting bone resorption and tooth movement, using a new orthodontic model in mice in which maxillary molars are moved for prolonged periods by near-constant, clinically relevant forces. Osteoclast influx to compression sites was initiated on day 3, was maximal on day 4, and persisted until at least day 12 after force application. Tooth movement paralleled osteoclast numbers. Minimal osteoclast apoptosis was observed, suggesting that recruitment, rather than programmed cell death, is a critical regulatory mechanism under conditions of constant force. Osteoclasts were reduced at compression sites by both OPG (95%) and pamidronate (70%); tooth movement was more dramatically inhibited by OPG (77% vs. 34%). Our findings indicate that constant orthodontic force regulates the recruitment, activation, and viability of osteoclasts, and that OPG could have clinical utility in preventing undesired tooth movement.