DataSheet_1_Real-world treatment patterns and outcomes among individuals receiving first-line pembrolizumab therapy for recurrent/metastatic head and neck squamous cell carcinoma.docx

BackgroundPembrolizumab, a PD-1 immune checkpoint inhibitor, is approved as first-line (1L) treatment for recurrent or metastatic head and neck squamous cell carcinoma (R/M HNSCC) as monotherapy or in combination with platinum and 5-fluorouracil chemotherapy. Limited data exist on the use of these regimens in real-world settings.ObjectiveOur primary objectives were to describe baseline characteristics and real-world overall survival (rwOS), time on treatment (rwToT), and time to next treatment (rwTTNT) among individuals with R/M HNSCC receiving approved 1L pembrolizumab therapies. We also aimed to identify baseline factors associated with choice of 1L pembrolizumab therapy and with rwOS.MethodsThis was a retrospective cohort study of adults with R/M HNSCC receiving 1L pembrolizumab monotherapy or pembrolizumab plus chemotherapy. We used Kaplan-Meier analyses to assess real-world outcomes, logistic regression modeling to identify factors associated with choice of 1L pembrolizumab therapy, and Cox proportional hazards models to identify factors associated with rwOS.ResultsThe study population included 431 individuals receiving 1L pembrolizumab monotherapy and 215 receiving 1L pembrolizumab plus chemotherapy. The use of 1L pembrolizumab monotherapy was associated with higher baseline combined positive score for PD-L1 expression, older age, higher Eastern Cooperative Oncology Group performance status (ECOG PS), laryngeal tumor site, and human papillomavirus (HPV)-positive tumor status. The pembrolizumab monotherapy group had a median (95% CI) rwOS of 12.1 (9.2–15.1) months, rwToT of 4.2 (3.5–4.6) months, and rwTTNT of 6.5 (5.4–7.4) months. Among this group, HPV-positive tumor status and lower ECOG PS were associated with longer rwOS, and oral cavity tumor site with shorter rwOS. The pembrolizumab plus chemotherapy cohort had a median (95% CI) rwOS of 11.9 (9.0–16.0) months, rwToT of 4.9 (3.8–5.6) months, and rwTTNT of 6.6 (5.8–8.3) months. In this group, HPV-positive tumor status was associated with longer rwOS.ConclusionsThis study adds to clinical trial data by summarizing real-world treatment outcomes with 1L pembrolizumab-containing therapies in a more heterogeneous population. Overall survival outcomes in both treatment groups were similar to those observed in the registration clinical trial. These findings support the use of pembrolizumab as standard of care for R/M HNSCC.</p

α-GalCer suppressed the production of autoantibodies against GPI, but not serum inflammatory cytokines.

<p>Mice were immunized with GPI peptide and treated with either DMSO or α-GalCer. Serum samples were obtained on days 1, 3, 6, and 28 after the immunization. (<b>A</b>) Levels of anti-recombinant human GPI antibody (total IgG, IgG1, IgG2a, IgG2b, IgG3) in sera obtained on day 28 were measured by ELISA (n = 11). Levels of (<b>B</b>) TNF-α and (<b>C</b>) IL-6 in the sera were measured by CBA (n = 3–5). Data are mean±SD. *p<0.05, by Man-Whitney analysis.</p

α-GalCer does not induce expansion of CD25+foxp3+ regulatory T (Treg) cells.

<p>Mice were immunized with GPI peptide and treated with either DMSO or α-GalCer, then euthanized on day 6 or 10 and dLN cells were harvested. (<b>A</b>) Foxp3+ T reg cells were detected as TCRβ, foxp3- and CD25-positive cells by flow cytometry. (<b>B</b>) Proportion of foxp3+ CD25+ cells among TCRβ+ αβ T cells. Data are mean±SD. *p<0.05, by Man-Whitney analysis.</p

iNKT cells do not directly suppress antigen-specific CD4+ T cells <i>in vitro</i>.

<p>Mice were immunized with GPI peptide and treated with either DMSO or α-GalCer. Mice were euthanized on day 10 and dLNs were harvested. (<b>A</b>) Total CD4+T cells (CD3+ CD4+ cells) and NKT-depleted CD4+T cells (CD3+ CD4+ α-GalCer loaded CD1d-tetramer negative cells) were sorted by flow cytometry. These cells were cultured with sorted MMC-treated CD3-negative cells as APC in the presence of 10 µM of GPI peptide for 24 h. (<b>B</b>) Supernatants were collected and subjects to quantitative analysis of IL-17 and IFN-γ levels by ELISA. Data are mean±SD. (n = 4–5). *p<0.05, **p<0.01, by Man-Whitney analysis.</p

Expansion of iNKT cells in draining lymph nodes of α-GalCer-treated mice.

<p>Mice were immunized with GPI peptide and then treated with either DMSO or α-GalCer and euthanized on day 6 or 10. The dLNs were obtained and examined for the presence of iNKT cells by FCM (Data are mean±SD. n = 4–5. Data are representative results of two experiments). (<b>A</b>) iNKT cells were detected as TCRβ and α-GalCer-loaded CD1d-tetramer double positive cells. (<b>B</b>) Proportion of iNKT cells among TCRβ+ αβT cells, and (<b>C</b>) absolute number of iNKT cells in dLNs of naive or immunized mice. *p<0.05 by Man-Whitney analysis.</p

α-GalCer significantly suppressed the severity of GPI peptide-induced arthritis.

<p>DBA1 mice were immunized with GPI peptide and then treated with either DMSO (as a vehicle control) or α-GalCer followed by clinical assessment of arthritis. (n = 5, data shown are representative results of three experiments). (<b>A</b>) Severe swelling of ankle joints of control mice and markedly improved swelling of the ankle joints of α-GalCer-treated mice. (<b>B</b>) Clinical score, and (<b>C</b>) Ankle thickness. The latter represented the severity of arthritis. Arrow indicates α-GalCer administration. Data are mean±SD. *p<0.01, by Man-Whitney test.</p

Suppression of antigen-specific CD4+ T cells by α-GalCer.

<p>(<b>A</b>)(<b>B</b>) Mice were immunized with GPI peptide and treated with either DMSO or α-GalCer. On day 6 (A) or day 10 (B), mice were euthanized and dLNs and spleens were harvested. CD4+ T cells were isolated from dLNs using MACS and cultured with mitomycin-treated splenocytes as antigen-presenting cells (APC). After 24-h culture, the levels of IL-17, IFN-γ, IL-2, TNF-α, IL-4 and IL-10 in the supernatant were measured by ELISA (n = 4). (<b>C</b>) Ten days after immunization, the mice were euthanized and dLNs and spleens were harvested. CD4+ T cells isolated from control or α-GalCer-treated mice were cultured with splenocytes (described as APC in the figure) from control or α-GalCer-treated mice. Cytokine levels in the culture supernatants were measured by ELISA (n = 4). Data are mean±SD. *p<0.05, by Man-Whitney analysis.</p

DataSheet_1_Real-world use of first-line pembrolizumab + platinum + taxane combination regimens in recurrent / metastatic head and neck squamous cell carcinoma.docx

IntroductionThe programmed death-1 (PD-1) immune checkpoint inhibitor pembrolizumab is currently approved in the US for the first-line (1L) treatment of recurrent or metastatic head and neck squamous cell carcinoma (R/M HNSCC), either alone or in combination with platinum and 5-fluorouracil (5-FU). However, the toxicity of 5-FU has motivated the study of alternate combinations that replace 5-FU with a taxane. The objective of the current study was to describe the baseline characteristics, treatment patterns and sequences, and real-world outcomes of individuals receiving pembrolizumab + platinum + taxane as 1L treatment for R/M HNSCC in the US.MethodsThis was a retrospective study of US adults ≥18 years of age receiving pembrolizumab + platinum + taxane as 1L treatment for R/M HNSCC, using electronic health record data from a nationwide de-identified database. Real-world overall survival (rwOS), time on treatment (rwToT), and time to next treatment (rwTTNT) outcomes were assessed using Kaplan–Meier analysis.ResultsThe study population comprised 83 individuals (80.7% male) with a median age of 64 years. The most common tumor site was the oropharynx (48.2%); 70.0% of these tumors were HPV-positive. A total of 71.1% of the study population had an Eastern Cooperative Oncology Group performance status of 0–1 at index date, 71.8% had a combined positive score for programmed death ligand-1 (PD-L1) expression of ≥1, and 30.8% had a score of ≥20. The median (95% CI) rwOS was 14.9 (8.8–23.3) months, rwToT was 5.3 (4.0–8.2) months, and rwTTNT was 8.7 (6.8–12.3) months. Among the 24 individuals who received a subsequent therapy, the most common second-line therapies were cetuximab-based (n = 9) or pembrolizumab-containing (n = 8) regimens.ConclusionsThe rwOS and other real-world outcomes observed for this study population further support pembrolizumab + platinum + taxane combination therapy as a potential 1L treatment option for R/M HNSCC.</p

Therapeutic effect of anti-TNF mAb, CTLA-4Ig, and anti-IL-6 mAb in GPI-induced arthritis

Glucose-6-phosphate isomerase (GPI)-immunized mice were treated with anti-tumor necrosis factor (TNF)-α mAb; anti-IFN-γ mAb or anti-IL-12 mAb; cytotoxic T-lymphocyte antigen 4 immunoglobulin (CTLA-4Ig; on days 8 and 12); CTLA-4Ig (on days 12 and 16); and or anti-IL-6 mAb just after the onset of arthritis (on day 8, on days 8 and 12, or days 12 and 16; arrow). The mean clinical index (± standard error) was examined throughout the study. *< 0.05, **< 0.01, by Mann-Whitney's U test. n = 6 mice in each group. Hematoxylin and eosin staining at day 21 (×40) is also shown: anti-TNF-α mAb, control antibody, and CTLA-4Ig (on days 8 and 12).<p><b>Copyright information:</b></p><p>Taken from "Therapeutic effects of antibodies to tumor necrosis factor-α, interleukin-6 and cytotoxic T-lymphocyte antigen 4 immunoglobulin in mice with glucose-6-phosphate isomerase induced arthritis"</p><p>http://arthritis-research.com/content/10/3/R66</p><p>Arthritis Research & Therapy 2008;10(3):R66-R66.</p><p>Published online 5 Jun 2008</p><p>PMCID:PMC2483457.</p><p></p

Concentration of inflammatory cytokines in serum of mice with GPI-induced arthritis

After immunization with glucose-6-phosphate isomerase (GPI), serum samples were collected from GPI-induced DBA/1 mice at the indicated time points (days 0, 7, 14, and 28). Serum concentrations of tumor necrosis factor (TNF)-α (solid circle), IL-6 (open square), IL-1β (open triangle), or IFN-γ (open diamond) were determined by enzyme-linked immunosorbent assay. Data are expressed as mean ± standard error. = 3 mice in each group. *< 0.05, by Mann-Whitney's U-test.<p><b>Copyright information:</b></p><p>Taken from "Therapeutic effects of antibodies to tumor necrosis factor-α, interleukin-6 and cytotoxic T-lymphocyte antigen 4 immunoglobulin in mice with glucose-6-phosphate isomerase induced arthritis"</p><p>http://arthritis-research.com/content/10/3/R66</p><p>Arthritis Research & Therapy 2008;10(3):R66-R66.</p><p>Published online 5 Jun 2008</p><p>PMCID:PMC2483457.</p><p></p