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Supplemental Tables 1-6 Supplemental Table 1. Results of some recent trials of chemotherapy in MPM. Supplemental Table 2. Post-Trial Therapies Supplemental Table 3. Characteristics of Patients Selected for Flow Cytometry Analysis Supplemental Table 4. List of Immune Response Genes assayed by Nanostring Supplemental Table 5: Correlation of Antibody Response to MOS or Radiographic Response Supplemental Table 6. Flow Cytometry Results

Purpose: “In situ vaccination” using immunogene therapy has the ability to induce polyclonal antitumor responses directed by the patient's immune system.Experimental Design: Patients with unresectable malignant pleural mesothelioma (MPM) received two intrapleural doses of a replication-defective adenoviral vector containing the human IFNα2b gene (Ad.IFN) concomitant with a 14-day course of celecoxib followed by chemotherapy. Primary outcomes were safety, toxicity, and objective response rate; secondary outcomes included progression-free and overall survival. Biocorrelates on blood and tumor were measured.Results: Forty subjects were treated: 18 received first-line pemetrexed-based chemotherapy, 22 received second-line chemotherapy with pemetrexed (n = 7) or gemcitabine (n = 15). Treatment was generally well tolerated. The overall response rate was 25%, and the disease control rate was 88%. Median overall survival (MOS) for all patients with epithelial histology was 21 months versus 7 months for patients with nonepithelial histology. MOS in the first-line cohort was 12.5 months, whereas MOS for the second-line cohort was 21.5 months, with 32% of patients alive at 2 years. No biologic parameters were found to correlate with response, including numbers of activated blood T cells or NK cells, regulatory T cells in blood, peak levels of IFNα in blood or pleural fluid, induction of antitumor antibodies, nor an immune-gene signature in pretreatment biopsies.Conclusions: The combination of intrapleural Ad.IFN, celecoxib, and chemotherapy proved safe in patients with MPM. OS rate was significantly higher than historical controls in the second-line group. Results of this study support proceeding with a multicenter randomized clinical trial of chemo-immunogene therapy versus standard chemotherapy alone. Clin Cancer Res; 22(15); 3791–800. ©2016 AACR.

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Additional Trial related information and methods

Supplemental Figure 1. Immunoblots of Patient Serum before and after Therapy Supplemental Figure 2. Distribution of 1/Nab Titers for the 40 Patients Supplemental Figure 3. Distribution of the Day 2 serum levels of Interferon-alpha Supplemental Figure 4. Lack of Correlation of lymphocyte or macrophage infiltration with survival. Supplemental Figure 5. Lack of Correlation of PDL1 staining with survival. Supplemental Figure 6. Correlation of the mRNA immunoscore with survival.

In chronic lymphocytic leukemia (CLL) patients who achieve a complete remission (CR) to anti-CD19 chimeric antigen receptor T cells (CART-19), remissions are remarkably durable. Preclinical data suggesting synergy between CART-19 and the Bruton’s tyrosine kinase (BTK) inhibitor ibrutinib prompted us to conduct a prospective single-center phase 2 trial in which we added autologous anti-CD19 humanized binding domain T cells (huCART-19) to ibrutinib in patients with CLL not in CR despite ≥6 months of ibrutinib. The primary endpoints were safety, feasibility, and achievement of a CR within 3 months. Of 20 enrolled patients, 19 received huCART-19. The median follow-up for all infused patients was 41 months (range, 0.25-58 months). Eighteen patients developed cytokine release syndrome (CRS; grade 1-2 in 15 of 18 subjects), and 5 developed neurotoxicity (grade 1-2 in 4 patients, grade 4 in 1 patient). While the 3-month CR rate among International Working Group on CLL (iwCLL)-evaluable patients was 44% (90% confidence interval [CI], 23-67%), at 12 months, 72% of patients tested had no measurable residual disease (MRD). The estimated overall and progression-free survival at 48 months were 84% and 70%, respectively. Of 15 patients with undetectable MRD at 3 or 6 months, 13 remain in ongoing CR at the last follow-up. In patients with CLL not achieving a CR despite ≥6 months of ibrutinib, adding huCART-19 mediated a high rate of deep and durable remissions. ClinicalTrials.gov number, NCT02640209.

Patients with cancer are ideally screened for symptoms, including distress, using patient-reported outcome measures (PROMs). This initiative was developed to ensure patients without access to an electronic portal were screened for distress and related symptoms during the COVID-19 pandemic. Prior to the pandemic, these patients could complete screening in clinic. However, many visits transitioned to telehealth. We implemented a standardized telephone outreach process targeting patients without active electronic portal accounts to improve remote symptom monitoring. Outreach resulted in 172 completed screens, identifying 110 needs for 63 individuals. Twenty-eight patients completed patient portal enrollment. Outreach calls captured a higher percentage of Black patients (34%) and a higher percentage of 61-80 year olds (69%) compared to portal users. Telephone outreach during the pandemic captured data that otherwise would have been missed in elderly and minority patients without electronic patient portal access. Patient engagement is vital to the distress screening process.

Purpose: “In situ vaccination” using immunogene therapy has the ability to induce polyclonal antitumor responses directed by the patient's immune system. Experimental Design: Patients with unresectable malignant pleural mesothelioma (MPM) received two intrapleural doses of a replication-defective adenoviral vector containing the human IFNα2b gene (Ad.IFN) concomitant with a 14-day course of celecoxib followed by chemotherapy. Primary outcomes were safety, toxicity, and objective response rate; secondary outcomes included progression-free and overall survival. Biocorrelates on blood and tumor were measured. Results: Forty subjects were treated: 18 received first-line pemetrexed-based chemotherapy, 22 received second-line chemotherapy with pemetrexed (n = 7) or gemcitabine (n = 15). Treatment was generally well tolerated. The overall response rate was 25%, and the disease control rate was 88%. Median overall survival (MOS) for all patients with epithelial histology was 21 months versus 7 months for patients with nonepithelial histology. MOS in the first-line cohort was 12.5 months, whereas MOS for the second-line cohort was 21.5 months, with 32% of patients alive at 2 years. No biologic parameters were found to correlate with response, including numbers of activated blood T cells or NK cells, regulatory T cells in blood, peak levels of IFNα in blood or pleural fluid, induction of antitumor antibodies, nor an immune-gene signature in pretreatment biopsies. Conclusions: The combination of intrapleural Ad.IFN, celecoxib, and chemotherapy proved safe in patients with MPM. OS rate was significantly higher than historical controls in the second-line group. Results of this study support proceeding with a multicenter randomized clinical trial of chemo-immunogene therapy versus standard chemotherapy alone. Clin Cancer Res; 22(15); 3791–800. ©2016 AACR.

Gene-mediated cytotoxic immunotherapy (GMCI) is an immune strategy implemented through local delivery of an adenovirus-based vector expressing the thymidine kinase gene (aglatimagene besadenovec, AdV-tk) followed by anti-herpetic prodrug valacyclovir. A phase I dose escalation trial of GMCI followed by chemotherapy was conducted in patients with malignant pleural effusion (MPE). AdV-tk was administered intrapleurally (IP) in three cohorts at a dose of 1 × 10(12) to 10(13) vector particles. Primary endpoint was safety; secondary endpoints included response rate, progression-free survival, and overall survival. Nineteen patients were enrolled: median age 67 years; 14 with malignant mesothelioma, 4 non-small-cell lung cancer (NSCLC), and 1 breast cancer. There were no dose limiting toxicities. All 3 patients in cohort 2 experienced transient cytokine release syndrome (CRS). Addition of celecoxib in cohort 3 reduced the incidence and severity of CRS (none > grade 2). Three patients are alive (23–33 months after GMCI), and 3 of 4 NSCLC patients had prolonged disease stabilization; one is alive 29 months after GMCI, 3.6 years after initial diagnosis. GMCI was safe and well tolerated in combination with chemotherapy in patients with MPE and showed encouraging response. Further studies are warranted to determine efficacy.

Background Immunotherapy with anti-CD19 CART cells (CART19) induces complete remission (CR) in the minority of patients with CLL; however, these CRs tend to be durable (Porter Sci Tr Med 2015). Based on preclinical evidence of synergy, we combined anti-CD19 CAR T cells with ibrutinib to test the hypothesis that pre- and concurrent treatment enhances the CR rate. Methods This is a pilot trial of autologous anti-CD19 CAR T cells in adults with CLL/SLL who were not in CR despite at least 6 months of ibrutinib. T cells were lentivirally transduced to express a CAR comprising CD3z, 4-1BB, and humanized anti-CD19 scFv (CTL119). Pts underwent lymphodepleting chemotherapy up to 1 week before infusion, followed by planned infusion of 1-5x108 CTL119 cells dosed as 10%, 30% and 60% of the total planned dose over 3 days, with doses beyond dose#1 given only in the absence of fever or cytokine release syndrome (CRS). Results CTL119 manufacturing was successful for all pts. Twenty pts were enrolled and 19 were infused (one pt was not infused due to intercurrent large cell transformation and newly diagnosed adenocarcinoma). Of the 19, 15 were male, the median age was 62 (range 42-76); and 5 were on 1st line ibrutinib. Of the remaining 14, the median number of prior therapies was 2 (range 1-16), and 3 pts had received prior murine CART19 therapy (CTL019) without ibrutinib. Eleven pts had abnormalities of chromosome 17p or TP53. An additional 3 pts had abnormalities of chromosome 11q22 or ATM. All pts had marrow involvement (median 21% range 7-63%). For 9 pts who had enlarged nodes at baseline, the median cross-sectional area was 1471 mm2 (range 178-2220). All pts received at least 2 CTL119 doses; 14 patients received all 3 and 5 received 2 doses. The median number of CART cells given was 5.3x106/kg (range 2.0-7.5). Median peak CART cell number by qPCR was 90,990 copies/ug genomic DNA (range 965-210,556) and by flow cytometry was median 536 CART cell/ul blood (range, 0-3640). 18/19 (95%) pts experienced CRS, with a median Penn CRS Grade of 2. CRS was grade 1-2 and 3-4 in 15 and 3 pts, respectively. Two pts received tocilizumab. Of 5 pts with encephalopathy, 2 were CTCAE gr 1, 2 gr 2 and 1 gr 4. One pt died on D14 from a cardiac arrhythmia during severe neurotoxicity after resolution of CRS. There were 49 gr 3 and 22 gr 4 toxicities in total. As of 7/16/18, 18/19 pts are alive (95%) and 12 pts have been followed for at least 12 months. The median follow-up for the 18 surviving pts is 18.5 months (range 8-28). Per iwCLL criteria, at 3 months 14 pts were evaluable and their responses were CR (n=6), PR (n=4), SD (n=3), PD (n=1). Marrow responses at month 3 were available in 18 and showed a morphologic CR in 17 pts (94%); of these 15 also had no measurable residual disease (MRD) by 9-color flow cytometry. MRD was also assessed at 3 months by deep sequencing of the immunoglobulin heavy chain locus (limit of detection 1 B cell in 1x106 nucleated cells). 14/18 pts were MRD negative, and the remaining 4 had 3.36, 4.76, 1.79, and 0.48 log10 reduction of the leukemic clonotype relative to the baseline sample. LN biopsies from 2 pts 3 and 10 months after CTL119 confirmed absence of the CLL clonotypes in this compartment as well. At 12 months, 11 pts had evaluable marrows of which 10 (91%) were in morphologic CR and 1 showed morphologic relapse. Of the 10 in morphologic CR, three pts showed low MRD positivity (3.58, 2.34, 3.79 log10 reduction) and the rest remained MRD-ve. Of the 3 pts who had received murine CTL019 previously, 2 were in MRD+ve CR at 12 months and one was refractory to humanized CTL119. Six pts discontinued ibrutinib at a median 8 months (range 3-12) due to toxicity (n=2) or pt choice (n=4). Five pts remain MRD-ve at short followup. In total, 16/18 pts remain in morphologic and/or flow CR at last followup. Conclusion In patients not achieving CR despite at least 6 months of ibrutinib who were treated with humanized CART19, we found an iwCLL CR rate of 43% and a bone marrow remission rate of 94% including a 78% MRD negative response by deep sequencing. This compares favorably to prior CART19 cell studies in patients with progressive CLL (iwCLL CR rates of 21-29%). CRS was frequent but mild-moderate and did not commonly require anti-cytokine therapy. These results suggest that the combination of CTL119 with ibrutinib results in a high rate of sustained responses and high rates of MRD-ve marrow response in patients with CLL. This combination will be further tested in larger studies. Figure. Figure. Disclosures Gill: Carisma Therapeutics: Equity Ownership; Extellia: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis: Research Funding. Frey:Servier Consultancy: Consultancy; Novartis: Consultancy. Mato:Johnson & Johnson: Consultancy; Portola: Research Funding; Acerta: Research Funding; AstraZeneca: Consultancy; AbbVie: Consultancy, Research Funding; Pharmacyclics, an AbbVie Company: Consultancy, Research Funding; Regeneron: Research Funding; TG Therapeutics: Consultancy, Research Funding; Celgene: Consultancy; Medscape: Honoraria; Prime Oncology: Honoraria. Lacey:Novartis Pharmaceuticals Corporation: Patents & Royalties; Tmunity: Research Funding; Parker Foundation: Research Funding; Novartis Pharmaceuticals Corporation: Research Funding. Melenhorst:Novartis: Patents & Royalties, Research Funding; Incyte: Research Funding; Tmunity: Research Funding; Shanghai UNICAR Therapy, Inc: Consultancy; CASI Pharmaceuticals: Consultancy. Davis:Novartis Institutes for Biomedical Research, Inc.: Patents & Royalties. Schuster:Gilead: Membership on an entity's Board of Directors or advisory committees; Physician's Education Source, LLC: Honoraria; Novartis Pharmaceuticals Corporation: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck: Consultancy, Honoraria, Research Funding; Pfizer: Membership on an entity's Board of Directors or advisory committees; Nordic Nanovector: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Dava Oncology: Consultancy, Honoraria; Genentech: Honoraria, Research Funding; OncLive: Honoraria. Siegel:Novartis: Research Funding. Isaacs:Novartis: Employment. June:Immune Design: Membership on an entity's Board of Directors or advisory committees; Celldex: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis Pharmaceutical Corporation: Patents & Royalties, Research Funding; Tmunity Therapeutics: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding; Tmunity Therapeutics: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding; Immune Design: Membership on an entity's Board of Directors or advisory committees; Novartis Pharmaceutical Corporation: Patents & Royalties, Research Funding. Porter:Kite Pharma: Other: Advisory board; Genentech: Other: Spouse employment; Novartis: Other: Advisory board, Patents & Royalties, Research Funding.

7509 Background: Immunotherapy with anti-CD19 CART cells induces complete remission (CR) in the minority of patients with CLL, but where CRs occur they tend to be durable. Based on preclinical evidence of synergy, we combined anti-CD19 CAR T cells with ibrutinib to test the hypothesis that pre- and concurrent treatment would enhance the CR rate. Methods: This is a pilot trial of anti-CD19 CAR T cells in adults with CLL/SLL who were not in CR despite at least 6 months of ibrutinib. Pts must have failed at least 1 regimen before ibrutinib, unless they had del(17)(p13.1) or a TP53 mutation. T cells were lentivirally transduced to express a CAR comprising CD3z, 4-1BB, and humanized anti-CD19 scFv (CTL119). Pts were lymphodepleted 1 week before infusion. Ibrutinib was continued throughout the trial. Results: Manufacturing was successful in all pts. Ten pts (9M, 1F; ages 47-77; 0-12 regimens prior to ibrutinib) have been infused. All had abnormalities of TP53 or ATM and two pts had increasing BTK C481S clones. Median marrow CLL burden was 10% (range 10-50%). The median follow-up is 6 months (range 0.5-9). Cytokine release syndrome (CRS) developed in 9 pts; gr1 in 2, gr2 in 6 and gr3 in 1 pt. One pt developed gr4 tumor lysis syndrome. Treatment of CRS with the IL-6 receptor antagonist tocilizumab was not required. At 3 months, 8 evaluable pts had achieved an MRD-ve marrow CR (89%) by 9-color flow, and all remain in marrow CR at last F/U. There was modest residual splenomegaly in 3/5 patients, and adenopathy resolved in 4/6 subjects with progression in 1/6. MRD assessment by deep sequencing will be presented. Conclusions: We observed 89% MRD-ve marrow CR in pts with high-risk CLL using a well-tolerated combination of CART cells and ibrutinib. Longer follow-up will reveal the durability of these results and could support evaluation of a first-line combination approach in an attempt to obviate the need for chronic therapy. Clinical trial information: NCT02640209.

The flexibility for learners to learn begins with the teacher's style of learning and the levels of cognition utilized in the classroom. Teachers have the basic capability to learn and teach; however, they are not all able to learn and teach effectively in the same exact way. The current study sought to determine the learning style and cognitive level of instruction of central Ohio horticulture teachers. The Florida Taxonomy of Cognitive Behaviors (FTCB) and the Group Embedded Figures Test (GEFT) were used to assess the level of cognitive instruction (FTCB) and the learning style (GEFT). The results indicated that 44% of the teachers preferred the field dependent learning style and 56% preferred the field-independent learning style. Furthermore, 84% of the teaching occurred at the lower levels of cognition. The mean weighted cognitive score for the teachers was 23.03. The mean weighted score reflected a cognitive level of teaching concentrated near the cognitive level of translation. A moderate positive (r=.32) relationship was found between learning style and the weighted cognitive level of instruction. Correlation coefficients between GEFT scores and the seven levels of cognition ranged from a substantial negative association (r= -.53) to a moderate positive association (r= .41). Bloom, Madaus, and Hastings (1981) labeled education as a "process of change," a process in which students must be changed in some way through the instruction they receive. Thus, when teaching in the classroom, educators need to be cognizant of the contribution to the development of the affective, psychomotor, and cognitive domains of learning. Regarding the cognitive domain, the educational literature (Gall, 1970; Roberts, 1974) has suggested that the emphasis in schools has been teaching students facts even though teachers and curriculum designers attested to the importance of teaching students to think. Supporting Gall's (1970) and Roberts' (1974) findings, studies conducted in agricultural education at the secondary level found that teachers concerned themselves with the subject matter students learned, more so than the cognitive level of their instruction (Cano, 1988; Cano & Newcomb, 1990). If insufficient instruction occurs at the higher levels of cognition, then students are not graduated adept at problem solving, analysis, and evaluation (Newcomb & Trefz, 1987).

3081Background: GMCI is a tumor-specific immune-stimulator through local delivery of aglatimagene besadenovec, an adenovirus-based vector expressing the HSV-1 thymidine kinase gene (AdV-tk) followe...