bluuebunny/crossref_metadata_2025 huggingface dataset:
Epidemiologic characteristics of colon cancer in Asian Indian and Pakistani population living in the USA compared to US White population in SEER database from 1988 to 2002
Authors: Sehbai A. S. , Hossain A. , Kurian S. , Abraham J.
Published: March 2020

20112

Background: About 1.5% of the US population is South East Asian in origin and it is one of the rapidly growing groups. Colon cancer is the second most common in male and third most common cancer in females in Asian Indian and Pakistani population living in the USA. This is the first study examining the epidemiologic characteristics of colon cancer in this group. Methods: SEER cancer incidence public use database have reported cancer incidence in 13 SEER registries from 1973 to 2002. Since 1988, Indian/Pakistani populations were reported separately under race and ethnicity. Frequency sessions were performed and descriptive parameters were calculated regarding age, sex, stage, grade, tumor location and diagnostic methods used. Results: A total of 4,229 cases of cancer were reported from 1988 to 2002 in this group and 352 (9%) were colon cancers. The reported incidence of colon cancer in SEER database is increasing (11 in 1988 vs. 53 in 2002). Table shows the comparative studies of different variables of colon cancer between Indian/Pakistani and US White population reported in SEER database. Conclusions: Our study shows that the incidence of colon cancer in Asian Indian and Pakistani population under age 50 is substantially higher than the US White population and the predilection is towards the male sex. Our results suggest that this ethnic group warrants close screening and risk factor stratification for earlier detection of colon cancer as well as awareness and counseling in high-risk families.

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No significant financial relationships to disclose.
Final results of a phase I and pharmacokinetic study of STEALTH liposomal CKD-602 (S-CKD602) in patients with advanced solid tumors
Authors: Zamboni W. C. , Friedland D. M. , Ramalingam S. , Edwards R. P. , Stoller R. G. , Belani C. P. , Strychor S. , Ou Y. C. , Tonda M. E. , Ramanathan R. K.
Published: March 2020

2013

Background: S-CKD602 is a pegylated STEALTH liposomal formulation of CKD-602, a semi-synthetic analogue of camptothecin (CPT) and topoisomerase I inhibitor. Preclinical studies have shown that prolonged exposure to CPT achieves the greatest antitumor activity. STEALTH liposomes prolong the circulation time in plasma, achieve high and extended drug exposure in tumor, and provide a convenient dosing schedule. Methods: Patients (pts) were administered S-CKD602 IV over 1 h once every 3 wks. Modified Fibonacci escalation was used (3–6 pts/cohort), and dose levels ranged from 0.1–2.5 mg/m2. Serial plasma samples were obtained prior to administration to 96 h after administration, and on days 8 and 15 of cycle 1. Plasma samples were processed to measure concentrations of encapsulated (E), released (R), and sum total (E+R) CKD602 total (lactone + hydroxyl acid) by LC-MS/MS. Area under the plasma conc versus time curve (AUC0-∞) and t1/2 were estimated. Results: 45 pts (21 male) have been treated: median age 62 y (range: 33–79 y); ECOG status: 0 and 1 (43 pts), 2 (2 pts). The majority of reported adverse events were grade (G) 1 and 2. Frequent nonhematological toxicities were asthenia, nausea, diarrhea, vomiting, and anorexia. Dose-limiting toxicities of G3 mucositis occurred in 1/6 pts at 0.3 mg/m2, G3/4 bone marrow suppression in 2/3 pts at 2.5 mg/m2, and G3 febrile neutropenia in 1/6 pts at 2.1 mg/m2. The maximum tolerated dose was 2.1 mg/m2. Stable disease for ≥ 6 cycles occurred in 5 pts (hepatocellular carcinoma (CA), thyroid, prostate, 2 pts sarcoma). Partial responses occurred in 2 pts with ovarian CA (1.7 and 2.1 mg/m2). At 2.1 mg/m2, the mean ± SD AUC and t1/2 of sum total S-CKD602 were 44 ± 33 μg/mL·h and 18 ± 8 h, respectively. In all plasma samples, >90% of drug was encapsulated. Conclusions: S-CKD602 represents a promising new STEALTH liposomal CPT agent with manageable toxicity and promising antitumor activity. Phase II studies of S-CKD602 at 2.1 mg/m2 IV once every 3 wks are planned. Prolonged plasma exposure over 1 to 2 wks is consistent with STEALTH liposomes and provides extended exposure compared with non-liposomal CPT. (Supported by ALZA and NIH/NCCR/GCRC #5M01 RR 00056).

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Phase I and pharmacokinetics (PK) study of sequential paclitaxel and trabectedin every 2 weeks in patients with advanced solid tumors
Authors: Papadopoulos K. P. , Chu Q. , Patnaik A. , Mita M. M. , Cooper J. , Van Maanen R. , Lopez-Lazaro L. , Lebedinsky C. , Rowinsky E. K. , Tolcher A. W.
Published: March 2020

2029

Background: Trabectedin (ET-743) is a DNA minor-groove intercalating agent that blocks transcription factor activity, elicits G2/M cell cycle arrest, and induces apoptosis. Single agent activity has been demonstrated in soft tissue sarcoma (STS), breast, prostate and ovarian cancer. In preclinical studies sequential exposure of paclitaxel (P) followed by trabectedin (T) 24 hrs later resulted in synergy. The objectives of this phase I study were to determine the maximum tolerated dose (MTD) and recommended phase II dose (RD) of sequential P and T administered every 2 weeks in patients with advanced solid tumors. Methods: Escalating doses of P (80–120 mg/m2) over 1hr iv on day 1 and T ( 525–775 μg/m2) as 3 hrs iv day 2 every 2 weeks were administered. To evaluate drug:drug interactions, P was administered alone on day -7 in course 1 and PK studies performed courses 1 & 2. MTD was defined as ≥ 2/3 pts with dose limiting toxicity (DLT) in the 1st 2 courses. Results: 29 pts were enrolled and 27 were evaluable: median age was 48 yrs (19–82). M/F 16/13. Pts were treated over 5 dose levels: I 80/525 (n = 3), II 80/580 (n = 3), III 120/580 (n = 6), IV 120/650 (n = 11), V 120/775 (n = 4). A total of 213 courses of therapy were given, median number of course was 4 (range 1–28). There were 4 DLTs due to neutropenia delaying therapy > 1 week; one each at 120/580 (Grade 2) and 120/650 (Grade 4) and 2 at the MTD of 120/775 (Grade 4). Most common toxicities observed for P+T were neutropenia (24%), nausea (51%), vomiting (24%), transaminitis (23%), myalgia (24%) and alopecia (20%). One pt with PNET has an ongoing CR 16 months, one breast cancer pt (prior P failure) has PR 12 mo+ and 8 pts (6 STS) had SD > 3 mo (range 4–15 mo). There appear to be no marked drug:drug interactions for P and T. Preliminary non-compartmental PK results [geometric mean (CV%)] for T include t1/2 52.4 hr (43.6%); Cl l/hr/m2 31.6 (57.3%); Vss l/m2 1382 (69%) and for P are t1/2, 29.2 hr (39.1%); Cl l/hr/m2 15.3 (44.1%); Vss 213 l/m2 (72%). Conclusions: The recommended dose for this combination is paclitaxel 120 mg/m2 day 1 and trabectedin 650 μg/m2 day 2 administered every 2 weeks. Antitumor activity in STS and breast cancer, predictable PK parameters, and tolerable toxicity warrants further study of this combination.

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A dose-escalating and pharmacologic study of bortezomib in adult cancer patients with impaired renal function
Authors: Mulkerin D. , Remick S. , Ramanathan R. , Hamilton A. , Takimoto C. , Davies A. , Ivy P. , Karol M. , Kolesar J. , Wright J.
Published: March 2020

2032

Background: Bortezomib is a highly selective and reversible inhibitor of the proteasome with activity in multiple myeloma and other malignancies. Patients (pts) with renal impairment have been treated in previous trials, but there has not been a systematic investigation into the effects of renal dysfunction on dosing. Study objectives were to characterize the pharmacokinetic (PK) and pharmacodynamic (PD) profile of bortezomib, and to determine the maximum tolerated dose (MTD) in adults with advanced malignancy and renal insufficiency ranging in severity from mild to dialysis dependence. Methods: Fifty-one pts have received intravenous bortezomib at 0.7 mg/m2 up to the approved dose of 1.3 mg/m2 on days 1,4, 8, and 11 every 3 weeks. Pts were stratified by 24-hour creatinine clearance (CrCL) normalized to a body surface area of 1.73 m2 into five cohorts per the table . Doses were escalated in cohorts of three pts in groups B-E. Blood samples were assayed for bortezomib concentration, as well as the PD endpoint of 20S inhibition. Results: Escalation of bortezomib doses to 1.3 mg/m2 was well tolerated in all groups with CrCL ≥ 20 mL/min/1.73 m2. There has been only one instance of dose limiting toxicity (group C at 1.3 mg/m2) which did not prevent successful completion of this cohort. No patients discontinued therapy due to renal deterioration. Doses of 0.7 mg/m2 were tolerable in Group D patients (CrCL< 20 mL/min/1.73 m2 ). Five dialysis pts have been treated; 3 at 0.7 mg/m2, and 2 at 1.0 mg/m2. All tolerated therapy well, and accrual to Groups D and E continues. PK and PD assays are underway and the analysis will be reported in full. Conclusions: This study is the first comprehensive evaluation of bortezomib in pts with various degrees of renal insufficiency including dialysis dependence. Bortezemib at the approved dose of 1.3 mg/m2 on this schedule is well tolerated by pts with CrCL ≥ 20 mL/min/1.73 m2. Results of this trial will allow for dosing recommendations for bortezomib use in pts with renal insufficiency.

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Final results from a phase I trial of ixabepilone in patients with advanced malignancies and lymphoma
Authors: Aghajanian C. , O’Connor O. , Cohen M. , Peck R. , Burris H.
Published: March 2020

2039

Background: Ixabepilone is the first analog in a new class of antineoplastic agents, the epothilones, which stabilizes microtubules and induces apoptosis. Ixabepilone has shown clinical activity in a broad range of tumors. Methods: This Phase I trial was designed to establish the maximum tolerated dose (MTD), dose-limiting toxicity (DLT), efficacy, safety, pharmacokinetics and pharmacodynamics of ixabepilone when administered as a 1-hour infusion every 3 weeks to patients with advanced solid tumors or lymphoma. Eligible patients were aged ≥18 years with histologically/cytologically confirmed non-hematologic cancer, or a pathologic diagnosis of relapsed/primary refractory non-Hodgkin’s lymphoma (NHL) or relapsed/primary refractory mantle cell lymphoma, with ≤CTC Grade 1 neuropathy. Ixabepilone doses ranged from 7.5–65 mg/m2. Response was assessed every 6 weeks using RECIST. DLT was defined as Grade 4 neutropenia and/or febrile neutropenia, thrombocytopenia, ≥Grade 3 nausea/vomiting and non-hematologic toxicity, or treatment delay of >2 weeks due to delayed recovery. Results: Of 61 patients (median age 58, range 18–81), 75% had solid tumors; 25% had lymphoma. 98% and 67% of patients had received one or ≥ two prior chemotherapy regimens, respectively. The MTD of ixabepilone as a 1-hour infusion every 3 weeks was established as 50 mg/m2. The most common DLTs were neutropenia, myalgia, arthralgia and stomatitis/pharyngitis. A total of eight patients (13%) achieved a durable objective response. Complete responses were achieved in two patients with primary peritoneal cancer and NHL. A partial response was seen in six patients. The most common Grade 3/4 treatment-related adverse events (only observed at doses ≥40 mg/m2) were sensory neuropathy (13%), fatigue (13%), myalgia (10%), arthralgia (7%), nausea (5%), febrile neutropenia (5%) and neutropenia (5%). Recovery to baseline or ≤Grade 1 neuropathy occurred in some patients. Conclusions: The recommended dose of ixabepilone for the initiation of Phase II studies based on this study is 50 mg/m2 over 1 hour every 3 weeks. Ixabepilone demonstrates promising safety in patients with solid tumors or lymphoma who have failed standard therapy. Encouraging activity was reported in several tumor types.

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Ixabepilone given weekly in patients with advanced malignancies: Final efficacy and safety results of a phase I trial
Authors: Dickson N. , Peck R. , Wu C. , Burris H.
Published: March 2020

2040

Background: Ixabepilone is a potent tubulin-targeting agent. The first epothilone analog, ixabepilone, has been developed to optimize the characteristics of the naturally occurring epothilone B. Methods: This open-label, single-arm Phase I dose escalation study was designed to establish the maximum tolerated dose (MTD), dose-limiting toxicity (DLT), safety and a recommended dose of ixabepilone when administered as a weekly infusion to patients (pts) with solid tumors who had failed standard therapy. Eligible pts were aged ≥18 years and had histologically/cytologically confirmed solid tumor disease and an ECOG status of 0–2. Ixabepilone was administered weekly as a 30-minute infusion on a 21-day schedule. Due to neurotoxicity, infusion time was increased to 1 hour with a 1-week break allowed. Dosing ranged from 1–30 mg/m2. Tumor evaluations were performed every 6 or 8 weeks (21-day or 28-day schedule, respectively). Toxicity was evaluated continuously. Results: 34 pts were treated on the 21-day schedule (median age 55, range 30–73) and 52 on the 28-day schedule (median age 55, range 33–79). 88% of pts had received prior chemotherapy. Grade 3 fatigue was the DLT in 2 of the 4 pts treated with 30 mg/m2 ixabepilone. No DLTs were seen at doses ≤25 mg/m2 on the 21-day schedule, or at doses of 15, 20 or 25 mg/m2 on the 28-day schedule. The MTD was 25 mg/m2 for the 21-day schedule. Of 12 pts who received this dose and schedule, 7 (58%) experienced Grade 1/2 neuropathy, 1 (8%) developed Grade 3 sensory neuropathy and 3 (25%) experienced Grade 3 fatigue. Of 52 pts who received 20 mg/m2 on the 28-day schedule, 2 (13%) had Grade 3 neutropenia, 1 (7%) had Grade 3/4 sensory neuropathy and 4 (27%) had Grade 3 fatigue. Five pts (two pts [21-day schedule], three pts [28-day schedule]) with breast, colon, ovary, head and neck tumors achieved objective partial responses. Conclusions: The recommended doses of ixabepilone from this study are 25 mg/m2 (30-minute weekly infusion, 21-day schedule) and 20 mg/m2 (1-hour weekly infusion, 28-day schedule). Ixabepilone demonstrated efficacy and an acceptable safety profile in pts with a broad range of tumor types, several of whom had failed a taxane.

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Phase I study of the effects of renal impairment on the pharmacokinetic (PK) and safety of satraplatin in patients (Pts) with refractory non-hematologic cancer
Authors: Hong D. S. , Galsky M. , Chiorean E. , Mulkerian D. , Greene D. , Nathan F. E. , Petrone M. , Camacho L. H.
Published: March 2020

2044

Background: Satraplatin (S) is a novel oral platinum analog with demonstrated activity in the treatment of pts with platinum-sensitive malignancies. A worldwide, double-blind, placebo-controlled randomized trial evaluating S as 2nd line therapy for hormone refractory prostate cancer (HRPC) has recently completed enrollment. The current study wasis designed to understand the effect of varying degrees of renal impairiment on the safety and PK of satraplatin. Methods: The study includes 4 levels of renal dysfunction, and 8 pts/cohort: Group 1 (G1) = Normal Renal Controls; G2 = Mild renal impairment (CrCl 50–80 mL/min); G3 = Mod. impairment (CrCl 30-<50 ml/min); G4 = Severe impairment (CrCl <30 mL/min). S was administered orally at 80mg/m2/d on d1–5 every 35 days. Results: 21 pts (of a planned total of 32) have been enrolled (13M/8F), median age 63 (range 45–72). Tumor types: Bladder (6), Renal (3), Breast (2), Prostate (2), Colon (2), Other (6). Among 15 evaluable pts, the cohort distribution is G1: 6 pts, G2: 4, G3: 4, and G4: 1. Twenty-six cycles of S have been delivered, and 11 pts have completed at least 2 cycles of therapy. Hematologic toxicities during the first 2 cycles include grade (G) 3/4 neutropenia (0 pts), G 3/4 thrombocytopenia (1), and G 3/4 anemia (1). No significant cardiac, renal, hepatic, or neurologic toxicity has been observed. Nausea, vomiting, and diarrhea were mild to moderate, and controlled with oral therapy. Of 4 pts with evaluable disease, 1 has stable disease, and 3 have progressed. Conclusions: S is well tolerated in pts with varying degrees of renal dysfunction. Updated safety and PK data will be presented at the meeting.

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Novel monoclonal antibodies for measuring 5-fluorouracil concentrations in biological fluids
Authors: Salamone S. J. , Courtney J. B. , Stocker D. , Harney R. L.
Published: March 2020

2055

Background: Therapeutic drug monitoring of anticancer drugs has potential for improving treatment of individual patients. 5-Fluorouracil (5-FU) has been reported to exhibit high pharmacokinetic variability and in some cases severe toxicity. Pharmacokinetic guided dosing of 5-FU using physical methodologies has demonstrated clinical usefulness in decreasing toxicity while resulting in equivalent or improved outcomes. The current physical methods for measuring 5-FU require specially trained personnel, are expensive, time consuming and therefore not amenable for widespread use. However, an automated immunoassay would provide a rapid, simple and inexpensive method to routinely measure levels of 5-FU in patients. Methods: In order to develop monoclonal antibodies that were selective for 5-FU, immunogens were prepared by attaching the drug to keyhole limpet hemocyanin (KLH) or bovine thyroglobulin (BTG) via a hydrocarbon tether bound through the N-3 nitrogen. Mice were immunized and the resulting antibodies were screened by ELISA. The selection of antibodies was based on sensitivity and cross-reactivity characteristics. Results: These immunogens were successful in eliciting 20 highly selective monoclonal antibodies for 5-FU with immunoassay dose response curves in the clinical range of 10 ng/mL to 5000 ng/mL. The immunoassay was tested with a mixture of the various pyrimidine compounds at their respective endogenous levels and shown to have no impact on the measurement of 5-FU. The Table below lists typical cross-reactivity performance characteristics that are elicited by this type of immunogen. Conclusion: This library of monoclonal antibodies represents the first report of antibodies selective for 5-fluorouracil. These antibodies will be useful for the rapid and cost effective analysis of 5-FU in human samples, providing a tool for optimization of 5-FU levels by pharmacokinetic guided dosing.

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Effects of rhIL-7 administration in humans on in vivo expansion of naïve, memory and effector subsets of CD4+ & CD8+ T-cells
Authors: Sportes C. , Hakim F. , Krumlauf M. , Babb R. , Fleisher T. , Brown M. , Engel J. , Buffet R. , Mackall C. , Gress R.
Published: March 2020

2504

Background: IL-7 has a critical and non-redundant role in T-cell lymphopoiesis and peripheral T-cell homeostasis. IL-7 administration may prove clinically valuable in conditions of disease induced (HIV) or iatrogenic T-cell depletion and for modulation of vaccine immune responses. In the first phase I study in humans, recombinant human interleukin-7 (“CYT 99–007”, Cytheris Inc., Rockville, MD) was administered subcutaneously every other day for two weeks in adults with refractory malignancies at 3, 10, 30 and 60 μg/kg/dose. Biologic activity, defined as a 50% increase over baseline of peripheral blood CD3+ T-cells, was seen at and above the 10μg/kg/dose in all patients. The kinetics of proliferation and expansion of peripheral blood T-cell subsets were analyzed. Methods: Multicolor flow cytometry was performed at baseline, 1, 2 and 3 weeks. Among CD4+ cells, the most naïve were defined as CD45RA+ /CD31+. Among CD4+ & CD8+ cells, the main naïve, memory and effector populations were defined respectively as CD45RA+/CD27+, CD45RA-/CD27+ and CD45RA-/CD27-. Within each subset, the number of cells in cycle was defined by Ki67 staining. Results: Following IL-7 therapy, there was marked proliferation of all T-cells subsets, peaking at week 1, most striking for the naive subsets with 30–70% of circulating cells induced to cycle. Proliferation rates were halved by week 2 despite continuation of treatment, coincident with the observed down-regulation of the IL-7 receptor. Cycling returned to baseline by week 3. Significant proliferation was also induced in effector and memory CD4+ and CD8+ T-cells but to a lesser magnitude, resulting in a greater net expansion of the naïve subsets, still ongoing one week after the end of treatment. Conclusions: IL-7 administration induces marked expansion of naïve, memory and effector CD4+ & CD8+ T-cells in humans. Consistent with the known down-regulation of the IL-7 receptor upon IL-7 exposure, proliferation rates decrease during the second week of treatment. rhIL-7 induced T-cell expansion may prove clinically valuable in adoptive immunotherapy as an adjunct to tumor vaccination and / or immunorestorative agent.

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A phase I trial of combination immunotherapy with CTLA-4 blockade and GM-CSF in hormone-refractory prostate cancer
Authors: Fong L. , Kavanagh B. , Rini B. I. , Shaw V. , Weinberg V. , Small E. J.
Published: March 2020

2508

Background: CTLA-4 is an costimulatory molecule expressed on activated T cells that delivers an inhibitory signal to these T cells. CTLA-4 blockade with antibody treatment augments T cell responses and anti-tumor immunity in animal models. Clinical trials with anti-CTLA-4 antibody treatment have demonstrated clinical responses in different malignancies including melanoma and hormone-refractory prostate cancer (HRPC). We have also shown that administration of granulocyte-macrophage colony-stimulating factor (GM-CSF) can also induce PSA declines in HRPC patients, presumably through enhancing presentation of endogenous antigens. The current study examines whether combining systemic GM-CSF to CTLA-4 blockade can augment immune and/or clinical responses in HRPC patients. Methods: In a phase I trial of patients with metastatic HRPC, sequential cohorts of 3–6 patients received GM-CSF (sargramostim, Berlex) 250 mg/m2/d SC on days 1–14 of a 28-day cycle with escalating doses (0.5, 1.5 or 3 mg/kg) of ipilimumab (MDX-010), a fully human anti-CTLA antibody (Medarex/BMS), given IV on day 1 of each cycle x 4 cycles. Patients were monitored for toxicity as well as for T cell activation. PSA and radiographic tests were performed at baseline and through therapy to evaluate for clinical response. Results: 18 patients were accrued. Ipilimumab-related dose-limiting toxicity was limited to one patient with grade 3 rash at the 3 mg/kg priming dose level. Seven patients had <50% declines in their serum PSA levels. A dose response relationship was seen between ipilimumab dose and activation of both CD4 and CD8 T cells in the blood. These effects were increased compared to effects seen with ipilimumab treatment alone in prior studies. Interferon-gamma production and lytic activity were also enhanced in circulating antigen-specific CD8+ T cells by the combination. Conclusions: GM-CSF may enhance T cell activation induced by CTLA-4 blockade. With increasing doses of anti-CTLA-4, both CD4 and CD8 T cell activation can be detected in the blood, consistent with a dose-response relationship. Supported by the UCSF Prostate SPORE NIH P50 CA89520.

No significant financial relationships to disclose.
Phase I, multicenter trial of CNTO 328, an anti-interleukin(IL)-6 monoclonal antibody (mAb) in subjects with selected hematologic malignancies
Authors: Kurzrock R. , Voorhees P. , Fayad L. , Orlowski R. , Van Rhee F. , Furman R. , Borghaei H. , Zaki M. H. , Prabhakar U. , Garay C.
Published: March 2020

2513

Background: IL-6 is implicated in the pathophysiology of several malignant diseases, including multiple myeloma (MM) and other hematological malignancies. CNTO 328 is a chimeric monoclonal antibody (mAb) that specifically binds and neutralizes human IL-6 with high affinity. A phase I trial is being conducted to assess safety and pharmacokinetics (PK) of CNTO 328 in subjects with B-cell non-Hodgkin’s lymphoma (NHL), MM, or Castleman’s Disease (CD). Methods: This trial will enroll at least 6 patients (pts) onto each of 5 cohorts, with 3 pts having a baseline C-reactive protein (CRP) of <10 mg/L, and 3 with ≥10 mg/L. Dose limiting toxicity (DLT) is defined as a ≥ Grade 3 event (excluding hematologic toxicity), or ≥Grade 2 allergic reaction/hypersensitivity attributed to CNTO 328 after the first administration. Results: Results from the first 13 subjects (3 CD, 3 MM and 7 NHL) enrolled in Cohorts 1 and 2 are currently available. Pts have received a median number of 4 (range 2 to 12) intravenous doses. CNTO 328 alone was well tolerated without pre-medication, and no DLTs have been observed. Some grade 3 or 4 adverse events (AEs) such as nausea, vomiting, elevated triglycerides, and decreased neutrophils were observed. Two subjects experienced serious AEs (SAEs) that were deemed unrelated to CNTO 328. Each of 3 pts with CD showed a clinical response, including improvement of hemoglobin, reduction of fever, resolution of rash, or decrease in lymphadenopathy. In one case of mantle cell NHL there was a reduction in lymphadenopathy. Two of 4 pts with previously progressive MM showed stabilization of M-protein levels and in % plasma cells in the bone marrow after one cycle of therapy. PK data is being analyzed. Conclusions: Treatment with CNTO 328 has been well tolerated at doses up to 6 mg/kg q 2 weeks. No clinical immune reaction to this chimeric mAb has been observed, and early evidence of tumor response and disease control has been observed.

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Endothelin receptor B predicts poor prognosis in epithelial ovarian cancer (EOC) and can be inhibited to increase intratumoral T cells and enhance tumor vaccine therapy
Authors: Buckanovich R. J. , Kim S. , Benencia F. , Sasaroli D. , Gimotty P. , Coukos G.
Published: March 2020

2524

Background: An inability of T cells to penetrate into tumor islets leads to a poor clinical outcome. We sought to identify tumor endothelial regulators of T cell trafficking. Methods: We used immunohistochemistry-guided laser capture microdissection to isolate and profile of vascular cells from EOC with or without (12 each) intratumoral T cells (ITC). Quantitative PCR (qPCR) and western blot of 60 EOC tumors was used to confirm array results and quantitate expression. Kaplan-Meier curves were generated to estimate survival according to high and low expression of ETRB (as determined by ROC analysis). Cox proportional hazard model was used to estimate the hazard ratio (HR). ETRB peptide antagonist BQ-788 was given QOD for 2 weeks to anti-tumor vaccinated mice with established ovarian tumors. After 8 weeks we used FACS and IHC to assess the presence of tumor-infiltrating T cells. Results: We identified ETRB as upregulated by the tumor vascular cells from tumors lacking ITCs. qPCR showed ETRB mRNA was 5.4 fold increased in ITC(-) vs. ITC(+) tumors. Univariate analyses demonstrated that ETRB expression was an independent prognosticator for both OS and DFS in EOC; the five-year overall survival rate was 70% for low ETRB expression versus <5%for high ETRB expression (p < 0.05, HR 0.35 (95% CI 0.17–0.71)). Bivariate analysis indicated that high ETRB expression predicted suboptimal cytoreduction (p < 0.05). High expression of ETRB strongly correlated with ITC (-). ETRB blockade in a murine tumor vaccine model with peptide BQ-788 restricted tumor growth, reduced ascites, and increased survival compared to control animals. Analysis of BQ-788 treated tumors revealed a 5–15× increase in CD4+ and CD8+ intratumoral T cells. Conclusion: ETRB is an important new biomarker, for EOC patients, that correlates with ITC status and predicts both suboptimal cytoreduction and poor clinical outcome. Antagonism of ETRB in mice can restore an otherwise ineffective anti-tumor immune response. This suggests that ETRB inhibition may be a powerful adjuvant for tumor immunotherapy.

No significant financial relationships to disclose.
A new method of constructing CD20/CD22 bispecfic antibody fusion proteins with improved direct lymphoma cytotoxicity compared to rituximab
Authors: Chang C. , Losman M. , Loo M. , Qu Z. , Rossi E. A. , Goldenberg D. M.
Published: March 2020

2536

Background: Anti-CD20 and anti-CD22 monoclonal antibodies (MAbs) have been shown to have antitumor activity in non-Hodgkin’s lymphoma (NHL) patients. Since preclinical and clinical trials also suggested that combining CD20 and CD22 MAbs, which have different mechanisms of action, could improve antitumor activity without a commensurate increase in toxicity, we hypothesized that a therapeutic advantage may be achieved with bispecific MAbs that bind simultaneously to both CD20 and CD22. Methods: A new platform technology, termed the Dock and Lock method (DNL), was successfully applied to produce a trivalent bispecific antibody, named TF3, which comprises two recombinant Fab fragments of hA20 (humanized anti-CD20 MAb; IMMU-106) stably tethered to one recombinant Fab fragment of epratuzumab (humanized anti-CD22 MAb; IMMU-103) via the specific interaction between a dimerization-and-docking domain and an anchoring domain appended to hA20 and epratuzumab, respectively. The cytotoxicity of TF3 was evaluated by cell-based assays using NHL cell lines. Results: TF3 is stable in both human and mouse sera andexhibitsthe samebinding affinity as hA20 IgG or epratuzumab Fab by competitive ELISA. With a 3-day MTT assay, TF3 at 10 nM inhibited 50% and 60% growth of Daudi and Ramos cells, respectively. Further, the observed anti-proliferative activity increased synergistically to >90% in the presence of anti-IgM (0.1 μg/mL). Results from a cell counting assay also demonstrated the ability of TF3 at 1 μM to completely inhibit the growth of Daudi and the potency of TF3 at 1 nM was comparable to that of rituximab at 1 μM under the same experimental conditions, reflecting a 1000-fold enhancement. Conclusions: These findings, to be extended by ongoing in vivo studies, suggest that the new DNL platform technology for making bispecific antibody fusion proteins provided a CD20/CD22 binding protein that is significantly more potent than rituximab in an in vitro NHL direct cytotoxicity assay.

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A novel rationally designed agonist antibody fragment that functionally mimics thrombopoietin
Authors: Renshaw M. , Frederickson S. , Lin B. , Su X. , Wang Y. , Bowdish K.
Published: March 2020

2541

Background: Thrombocytopenia, or low platelet count, is a significant clinical problem associated with ITP, cancer chemotherapy, or other clinical settings. To avoid transfusions, agents that stimulate platelet production are in development. Clinical trials with recombinant versions of thrombopoietin (TPO), which stimulates platelet production in a lineage specific manner by binding to cMpl receptor (cMpl-R) on megakaryocytic progenitors, were demonstrated to increase platelet counts in humans. However, the possible generation of an anti-TPO immune response that cross-reacts with and impairs the function of the endogenous cytokine is a significant disadvantage of this treatment approach. Alternate thrombopoietic agents that lack native TPO primary sequences have been developed to address this concern. Methods: Using rational design, antibody fragments (Fabs) that mimic TPO were created. A peptide with cMpl-R binding capability was grafted into different CDRs of a fully human Fab scaffold. Functional presentation of the peptide was optimized using phage display and cell-based panning. Select antibodies and fragments containing two grafted peptides were assayed for their ability to stimulate cMpl-R in vitro. In vivo stimulation of platelet production was tested in normal mice injected daily for five days with either rhTPO (90 μg/kg) or Fab59 (0.2 mg/kg, 2 mg/kg or 5 mg/kg) or negative control Fab (5mg/kg). Results: Several candidates demonstrated agonist activity in an in vitro cMpl-R signaling reporter assay, including Fab59 which was estimated to be equipotent to TPO. In vivo, a rise in peripheral platelet counts comparable to rhTPO was seen with Fab59 at 2 mg/kg and 5 mg/kg. Serum antibodies generated in response to dosing with Fab59 did not cross-react with murine or human TPO. Conclusion: These rationally-designed mimetic Fabs may provide a therapeutic intervention for thrombocytopenia while avoiding the potential generation of neutralizing antibodies to endogenous TPO.

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Immune evasion by melanoma and ovarian tumor cells through upregulation of the immunosuppressive molecule CD200
Authors: Siva A. , Xin H. , Qin F. , Mickel A. , Faas S. , Kretz-Rommel A. , Bowdish K. S.
Published: March 2020

2545

Background: Immune escape by tumors can occur by multiple mechanisms, each a significant barrier to immunotherapy. Upregulation of the immunosuppressive molecule CD200 on chronic lymphocytic leukemia cells inhibits Th1 cytokine production required for an effective cytotoxic T cell response. CD200 expression on human tumor cells in animal models prevents human lymphocytes from rejecting the tumor; treatment with an antagonistic anti-CD200 antibody restored lymphocyte-mediated tumor growth inhibition. This study evaluated CD200 expression on other cancers, and its effect on immune response. Methods: CD200 levels in ovarian adenocarcinoma and metastatic melanoma samples were evaluated by RT-QPCR and immunohistochemistry. Cell-surface CD200 on melanoma and ovarian cancer cell lines was assessed by flow cytometry. The effect of CD200 on cytokine production in mixed lymphocyte reactions (MLR) was assessed by adding the cells to cultures containing human monocyte-derived dendritic cells and allogeneic T cells. Th1 and Th2 cytokines in culture supernatants were detected by ELISA. Results: RT-QPCR showed CD200 expression levels upregulated in serous ovarian adenocarcinoma compared to normal samples. In malignant melanoma, CD200 expression in jejunum metastases was significantly higher than in normal samples, and 2 of 6 lung metastases showed CD200 upregulation. IHC showed strong, membrane-associated CD200 staining on malignant cells of two melanoma patients. Three ovarian cancer patients showed varying levels of CD200 tumor staining; all showed strong stromal staining. CD200 was highly expressed on the cell surface of SK-MEL-24 and SK-MEL-28 melanoma and OV-CAR-3 ovarian cancer cell lines and moderately expressed on the melanoma cell line SK-MEL-5. Addition of these cell lines to MLRs downregulated the production of Th1 cytokines; addition of CD200-negative cell lines did not. Inclusion of an antagonistic anti-CD200 antibody during the culture restored Th1 cytokine responses. Conclusion: Melanoma and ovarian tumor cells can upregulate CD200, thereby potentially suppressing anti-tumor immune responses. Therapy with an antagonistic anti-CD200 antibody may permit an effective cytotoxic immune response against the tumor cells.

[Table: see text]
Ex vivo activation and expansion of γδ T cells for immunotherapy of glioblastoma
Authors: Suarez-Cuervo C. , Bryant N. L. , Lopez R. D. , Gillespie G. Y. , Lamb L. S.
Published: March 2020

2553

Background: Activated γδ T cells will efficiently kill established glioblastoma (GBM) cell lines in vitro. However, their potential for cell therapy has been limited by their relatively small numbers in the peripheral blood and their sensitivity to activation induced cell death (AICD) during ex vivo expansion. Our laboratory developed a method whereby γδ T cells, when stimulated by IFN-γ, IL- 12, and anti-CD2, acquire resistance to AICD and can be expanded in culture with anti-CD3 and IL-2. The γδ T cells expanded by this method are highly cytotoxic to GBM. We have now sought to adapt the laboratory method for human use. Methods: The laboratory method and selection procedure were modified to employ pharmaceutical-grade reagents. Peripheral blood mononuclear cells (1.0 × 106/ml) were stimulated overnight with media containing cGMP grade human serum, IFN-γ, IL-12, and anti-CD2. Media containing human serum, IL-2 and anti-CD3 was then added (1 v/v) and refreshed 1 v/v 3× weekly. The cultures were harvested after two weeks followed by depletion of CD4+ and CD8+ T cells, thereby enriching γδ T cells. Cytotoxicity of expanded of γδ T cells was evaluated against GBM primary tumor cultures, established GBM cell lines, and cultured astrocytes using a commercial flow cytometric method. Results: After two weeks in culture, γδ T cells expanded up to 1000 fold (usually 200–400 fold), consistent with results from our previous research-scale method. Positive selection yielded a γδ T cell product with >80% purity and >70% recovery. The cell products exhibited incremental cytotoxicity against several primary GBM cultures and established GBM cell lines. Astrocytoma cells were not killed. Conclusions: Apoptosis-resistant γδ T cells can be expanded and selected using clinically approvable reagents and in numbers sufficient for immunotherapy of malignant brain tumors. Initial data show that expanded γδ T cells retain cytotoxicity against the GBM primary cultures and spare normal astrocytes.

No significant financial relationships to disclose.
Tumor stem cell vaccine for NSCLC and GBM-assessment of feasibility, adverse events and efficacy
Authors: Moviglia G. A. , Varela G. , Gaetta C. A. , Guerra M. , Carrizo A. , Costanzo H. , Farina P. , Bastos F. , Moviglia-Brandolino M. T. , Paes de Lima A. , Merino S.
Published: March 2020

2556

Background: Only a small part of the total cell population of a malignancy has the potential to grow and regenerate the total tumor mass. The rest are only differentiated forms of those stem cells. This biologic characteristic seems to be responsible for the failure to achieve long term remission for several peptide based vaccines. Method: To overcome this obstacle, we have established a technique that uses tumor stem cells (TSC) to develop a TSC-B cell vaccine. To test efficacy and best administration form of this vaccine, we have proceeded as follow: First: Pieces from total or partial resections of GBM or NSCLC were mechanically dissociated to a single cell suspension. Cells were cultured in an appropriate tissue culture medium and allowed to grow for two weeks or until they reach a semi confluent stage. Then, cells were harvested and re-seeded. This procedure was repeated 6 to 24 weeks according to vitality and growth characteristics of cells. Finally, Tumor cells were fused with autologous B cells. The hybrid was kept in culture and used for an intra lymph node immunization every 3 weeks. To potentiate its action, 3 days prior to its administration we co-incubated the hybrid with either autologous dendritic cells or autologous mononuclear cells. Adverse events were evaluated by the “Common terminology criteria for Adverse Events (AE), 2004” developed by NIH. Definitions are available on the web at http://ctep.cancer.gov/reporting/ctc.html . Clinical efficacy of this procedure was evaluated according RESIST criteria for NSCLC and a combination of MRI and spectroscopy for GBM. Results: At present 8 patients with GBM with post surgical second relapse and who had received complete radiotherapy doses were treated. We have also treated 6 patients with NSCLC:4 with minimal residual disease and 2 with measurable residual disease. Patients have presented minor adverse events, mainly referred to inflammation of the injection site. The median survival rates registered to the present are 22 months for GBM patients and 3 years for patients with stage IV NSCLC who had minimal residual disease at the beginning of the treatment. Conclusion: A tumor stem cell vaccine seems to be a feasible and effective therapy with very low toxicity for the treatment of advanced GBM and NSCLC patients.

No significant financial relationships to disclose.
Pharmacokinetics (PK) and immunologic responses in a phase I/II study of a sustained release formulation of IL-2 in renal cell carcinoma (RCC) patients
Authors: Oudard S. , Thiounn N. , Fridman H. , Amsellem-Ouazana D. , Colombel M. , Jacqmin D. , Malavaud B. , Kravtzoff R.
Published: March 2020

2558

Background: The utility of interleukin-2 (IL-2) in treating RCC is limited by its significant toxicity, expense, and limited efficacy. Formulations that reduce post-injection peak concentration (Cmax) and support sustained therapeutic levels may improve efficacy and tolerability of IL-2 therapy. In preclinical studies, IL-2 formulated with Medusa (IL-2XL), a novel sustained drug delivery system, reduces Cmax and supports controlled release of IL-2 for up to 6 days. Methods: The open-label Phase I/II study was designed to accrue 12 patients (pts), with a goal of 10 evaluable pts. Pts aged 18–85 at high risk of recurrence after surgical resection of histologically confirmed RCC, and without CNS metastases, other malignancy, autoimmune disorders were eligible. Pts received one single subcutaneous (SC) injection of 10.6 MIU naked IL-2 (IL-2N) or IL-2XL. At least two weeks later, patients were crossed over to receive a single SC administration of 10.6 MIU of the other agent. Safety, PK, biological activity and immunological responses were assessed. Results: 8 patients have accrued and results from 6 pts evaluable to date show a 4-fold reduction in Cmax following IL-2XL injection compared with IL-2N. IL-2 was detectable in serum 7 days after IL-2XL treatment, compared with 2 days for IL-2N. Relative bioavailability of IL-2XL was approximately 150% of IL-2N, and Tmax was appoximately 48 hrs compared with 2–4 hrs. T50 also was higher following IL-2XL administration compared with IL-2N (48 vs. 8 hrs). Soluble CD25 and neopterine, markers of IL-2 activity, had much higher Cmax (159 vs. 63 pg/ml and 14 vs. 6 pM for sCD25 and neopterin) and AUC (14,749 vs. 7,701 pg/ml/hrs and 1,569 vs. 665 pM/hrs for sCD25 and neopterin) following IL-2XL treatment compared with IL-2N. Compared to IL-2N immunoflowcytometry showed an increase in percentage and activation of lymphocytes (CD3+/CD45+; CD8+/CD25+ and CD4+/CD25+). Conclusions: Weekly dosing of IL-2XL reduces Cmax, increases induction of IL-2 biomarkers and improves immunologic responses compared with IL-2N. Data support the development of IL-2-XL used on a weekly schedule for treating cancer and viral disease such as HIV.

No significant financial relationships to disclose.
A novel non-HLA-restricted cellular immune assay for monitoring patient (pt) response to targeted immunotherapeutics
Authors: Manson K. , Dolan K. E. , Marshall J. , Kaufman H. , Mazzara G. P. , Schuetz T. J.
Published: March 2020

2567

Background: Many therapeutic cancer vaccines are designed to elicit T-cell immunity. In vitro monitoring of pt response is limited by the HLA-restriction of ELISPOT assays. We report a novel, non-restricted assay for measuring immune responses generated by PANVAC-VF, which comprises 2 recombinant viral vectors in a heterologous prime-boost immunization regimen. Each vector contains genes encoding the tumor antigens CEA and MUC-1, and 3 immunologic costimulatory molecules B7.1, LFA-3, and ICAM-1. The safety of PANVAC-VF was tested in a Phase 1 study of 10 pts with advanced pancreatic cancer (ASCO 2005 abstr 2576). Methods: Peripheral blood mononuclear cells (PBMCs) were drawn at baseline and each subsequent timepoint (∼2 wks after each injection) to measure cellular immune responses. PBMCs were separated from whole blood over a Ficoll gradient; isolated PBMCs were frozen in liquid nitrogen until analysis. A monkey breast cancer cell line (CMMT 110/C1) was infected with PANVAC-F or control fowlpox recombinant. On Day 2, PBMCs were incubated 10:1 with infected CMMT 110/C1 cells, and the mixtures co-cultivated for 72 h. Supernatants were assayed for interferon γ production. Results: Of 8 pts with PBMCs sufficient for testing, 5 developed an antigen-specific immune response to CEA and/or MUC-1. In general, interferon γ production steadily increased with repeat boosting immunizations. Cellular depletion experiments suggest that the result is dependent on CD4+ cells, allowing analysis of samples from all pts, unlike the ELISPOT, which is restricted to HLA-A2 peptide presentation. Preliminary analysis suggests a positive correlation of immune response with overall survival. Conclusions: We developed a novel, non-restricted cellular assay to assess immune response to a targeted immunotherapeutic, PANVAC-VF. Five of 8 assayed pts developed an antigen-specific cellular immune response. Pt numbers are small, but there appears to be a correlation between immune response and overall survival. This assay will be used for samples from a randomized Phase 3 study in pancreatic cancer and a randomized Phase 2 study in prostate cancer, generating a large data set of immune responses from all pts.

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Antitumor activity of a synthetic agonist of TLR9 in preclinical lung cancer models
Authors: Wang D. , Yu D. , Kandimalla E. R. , Agrawal S.
Published: March 2020

2568

Background: Synthetic agonists of TLR9 induce potent Th1-type innate and adaptive immune responses. In the present study, we have studied antitumor activity of a synthetic agonist of TLR9 referred to as immune modulatory oligonucleotide (IMO) in lung cancer models either alone or in combination with chemotherapeutic agents. Methods: Two different models are evaluated. In one model, mice implanted peritoneally with 3LL-C75 lung carcinoma cells were administered with IMO or PBS once in every two days for six times for evaluating antitumor activity of IMO alone. In a second model, to evaluate combination treatment of IMO with Gemcitabine or cyclophosphamide, mice bearing subcutaneously implanted 3LL tumor were administered with peritumoral injections of IMO and i.p. injections of Gemcitabine or cyclophosphamide. Results and Conclusions: Administration of IMO to mice bearing 3LL-C75 lung carcinoma inhibited tumor growth. Tumor free mice from this study failed to grow tumor when rechallenged with 3LL-C75 lung carcinoma cells, suggesting tumor bearing mice administered with IMO developed memory responses for the same tumor. Further more, naïve mice adoptively transferred with splenocytes obtained from mice that remained tumor free from the above treatment group failed to grow tumor to a rechallenge with 3LL-C75 tumor cells. The co-administration of IMO with chemotherapeutic agents, Gemcitabine and cyclophosphamide resulted in enhanced antitumor effects in 3LL lung cancer model. The present studies show potent antitumor activity of IMO when administered alone and in combination with Gemcitabine and cyclophosphamide in preclinical lung cancer models.

[Table: see text]