bluuebunny/crossref_metadata_2025 huggingface dataset:
Can gene expression microarray (GEM) provide a diagnosis in cases of carcinoma of unknown primary?
Authors: Bridgewater J. A. , Floore A. , Warmoes M. O. , Veer van’t L.
Published: March 2020

20117

Patients with CUP constitute approximately 5% of all malignancies and have no primary site of malignancy despite standard investigation, a poor prognosis secondary to metastatic disease and generate despondency amongst clinicians. GEM may provide an improved diagnostic tool and improve prognosis for patients through the use of more site specific chemotherapy. CupPrint is a 1900 gene custom microarray optimized for paraffin samples which uses a compiled database of 591 cancer cases to provide a statistically proven method for the discrimination of 39 different tumor types. Few data are available on the practical application of this novel technique. We have taken paraffin samples from 21 cases of “true” CUP where standard investigation failed to determine a primary site of malignancy although there were some cases were more uncertain than others. Three 4μm sections from the paraffin blocks were analysed and reviewed in the context of the histology reports, an independent histological review and the clinical history. 2 datasets were available for 1 patient and 3 datasets for another. A case by case analysis will be given but in summary, 16/21 diagnoses confirmed the clinicians suspicion. There was a clinical/CupPrint inconsistency in 4/21 patients and a pathological/CupPrint inconsistency in 1 patient. All CupPrint diagnoses were possible clinically. There was a consistency between identical samples analysed blindly by CupPrint and a consistency between different sites of tumour metastasis in the same patient. For most patients there was a consistency within the “K-Nearest Neighbors” analysis (see http://www.agendia.com). Clinical suspicion coincides with CupPrint certainty such that in those patients for whom there was no suspected site of primary, the “K-Nearest Neighbours” analysis did not match. 11/21 patients had had palliative chemotherapy with a further 4 yet to be receive treatment thus 15/21 patients could receive palliative chemotherapy. 5 patients are still alive. CupPrint is a promising analytical tool in order to more appropriately direct systemic palliative treatment in patients with CUP. These data need to be expanded to demonstrate a survival benefit of CupPrint guided treatment in a randomized clinical study.

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Phase I trial of a novel epothilone, KOS-1584, using a weekly dosing schedule
Authors: Stopeck A. , Thomas E. , Jones S. , Cohen J. , Cropp G. , Zhou Y. , Feng L. , Kersey K. , Hannah A. L. , Burris H.
Published: March 2020

2041

Background: Several epothilones are progressing through phase 1–3 clinical trials treating solid tumor malignancies. KOS-1584 is a 3rd generation epothilone with 3–12 fold increased potency compared to Epothilone D (as measured by in vitro cytotoxicity, in vivo xenografts or induction of G2/M arrest by flow cytometry) and improved pharmacologic/pharmacokinetic profile (enhanced tumor tissue penetration and reduced exposure to the CNS). This dose-escalation trial explores a weekly administration schedule of KOS-1584. Methods: Define the MTD, toxicity profile and PK of KOS-1584 when administered to patients with advanced solid malignancies via 1-hour infusion on Days 1, 8 and 15 every 4 weeks. Plasma PK and pharmacodynamics (serial sampling of PBMCs for soluble and polymerized microtubules by immunoblot) were assessed. Results: 12 pts (7 F; median age 60; median ECOG PS 1; median prior regimens 4, range 2–13) enrolled in 5 dose levels (0.8, 1.5, 2.5, 5.0 and 7.5 mg/m2). To date, no Cycle 1 DLT has been seen; one Grade 3 episode of arthritis occurred in Cycle 2. Drug-related toxicities, all Grade 1 or 2 in severity: fatigue (n=3), anorexia (n=2) and individual patients with constipation, nausea, mucositis, dehydration, headache and pruritus. Neurotoxicity has not been observed. PK/parent (n=8): t½ 18.5 ± 6.8h, Vz 504 ± 234 L and CL 19.7 ± 6.1 L/h (none of these parameters showed evidence of dose dependency). 5.0 mg/m2 Cmax 122.4 ± 60.6 ng/mL; AUCtot 688.2 ± 212 ng/mL*h. Dose proportional increase in exposure and Cmax was observed over the dose range tested to date. At 5.0 mg/m2 the1-hr infusion Cmax is 3-fold higher and AUCtot 50% higher than for the same dose delivered over 3 hours. As predicted by allometric scaling from animals, Vz is ∼4-fold and t½ 2-fold higher than that of Epothilone D. Activity consisted of extended stable disease (a patient with colon cancer for 3 cycles). Dose-dependent increases in polymerized microtubules were observed: prior to infusion ∼10% of tubulin was in its polymerized form; this increased to 20%, 30%, 35% and 48% for the 1st 4 dose levels at infusion end. Conclusions: Accrual is continuing in order to define the optimal dose on this regimen. Exposure and Cmax remain linear within this dose range; slower clearance is observed for the same dose administered over 1 hour compared to 3 hours.

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A finalized phase II report of pemetrexed (P) plus gemcitabine (G) as front-line chemotherapy for patients with peritoneal mesothelioma (PM)
Authors: Simon G. R. , Janne P. A. , Langer C. , Clarie V. , Dowlati A. , Gadgeel S. M. , Kelly K. , Ye Z. , Obasaju C. K. , Hood K. E. , Kindler H. L.
Published: March 2020

2050

Background: P in combination with cisplatin is approved for the treatment of malignant pleural mesothelioma. In an expanded access program, P ± cisplatin was shown to have activity in PM. P plus G are synergistic in preclinical models, but the activity of this combination in PM is unknown. The primary objective of this study was to determine time to disease progression in pts with PM receiving P plus G. Methods: Twenty pts were accrued. Treatment consisted of G 1250 mg/m2/30 minutes on D 1 and 8, and P 500mg/m2 /10 minutes on D 8, immediately before G. Treatment was repeated every 21 days for a total of 6 cycles or until progressive disease. All received folic acid, vitamin B12 and steroid prophylaxis. Results: Median age was 67.5 years (range: 46–79). Histology was epithelial:other = 14:6, M:F = 15:5, and ECOG PS 0:1:2 = 9:6:4. A total of 93 cycles (median 6) were administered. Patient-specific Grade 3/4 hematologic toxicity included neutropenia (60.0%), febrile neutopenia (10%; including one toxic death), anemia (5%), and thrombocytopenia (0%). Grade 3/4 nonhematologic events were fatigue (20.0%), constipation (10.0%), diarrhea (5.0%), peripheral neuropathy (5.0%), and dyspnea (5.0%). Median relative dose intensity was 94.5% for P and 82.9% for G respectively. Investigator-reported response rate was 15% (95% CI: 3.2–37.9%) with 1CR, 2PR, 7SD, 7PD, and 3 Unknown. Median time to progressive disease (MTTPD) and median survival (MS) were and 16.5 months (95% C: 6.6-TBD) and 19.1 months (95% CI: 6.6-TBD) respectively. Conclusions: P plus G is well tolerated in patients with PM. The clinical benefit rate (PR + SD) of 50%, MTTP and MS suggest that this non-platinum doublet can be a reasonable alternative to a platinum doublet in this disease.

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Pemetrexed in combination with paclitaxel: A phase I clinical and pharmacokinetic trial in patients with solid tumors
Authors: Graefe T. , Bolling C. , Lubbing C. , Latz J. , Blatter J. , Hanauske A.
Published: March 2020

2051

Background: Pemetrexed (Alimta [AL]) and paclitaxel (P) are clinically active in a variety of tumors. The primary objective of this trial was to determine the maximum tolerated dose (MTD) of the ALP combination; secondary objectives were: determination of dose-limiting toxicities (DLTs), definition of a recommended phase II dose, pharmacokinetic (PK) characterization and the anecdotal collection of antitumor activity. Methods: Escalating doses of P (3h infusion, d1 and d8) and AL (10 min infusion, d8 prior to P) were given in a 21d cycle. Results: 59 patients (pts) were enrolled. DLTs occurred at the following ALP (mg/m2) doses: 400/30 [G3 bilirubin (b), G3 and G4 thrombocytopenia (plts)]; 500/30 (G4 plts); 500/40 (G3 b); 500/75 (G4 ANC); 500/100 (G4 leukopenia, G4 ANC). With G4 leukopenia and G4 ANC in 4/6 pts and febrile neutropenia in 1 pt, the MTD was reached at the ALP (mg/m2) dose of 500/120. To confirm safety at the recommended dose-level, another 6 patients were treated at the ALP (mg/m2) dose of 500/100. 18 pts [mesothelioma (3), esophagus (2), lung (1), liver (1), renal (1), stomach (1), thyroid (9)] showed stable disease as best response. 4/14 (29%) pts with thyroid carcinoma showed long lasting partial responses [duration (months) 29+, 22, 18, 15]. One additional PR (2) was observed in a pt with penile carcinoma. AL PK when administered with P were consistent with those for AL administered as a single-agent. Conclusions: The ALP combination is safe and shows broad clinical activity. 500/100 mg/m2 is the recommended dose for further studies. Promising antitumor activity was observed in thyroid cancer. A phase II trial in thyroid carcinoma will be conducted.

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Dose escalation phase of a phase I/II study of GTI-2501, an antisense to the R1 subunit of ribonucleotide reductase (RNR) and docetaxel in patients with metastatic hormone-refractory prostate cancer
Authors: Ko Y. , North S. , Berry S. R. , Ernst D. S. , Klotz L. , Murray P. , Venner P.
Published: March 2020

2078

Background: GTI-2501 (GTI) is a 20-mer oligonucleotide that is complementary to the R1 subunit of the RNR mRNA. The R1 protein is overexpressed in multiple tumour cell lines. GTI displays anti-tumour activity against prostate cancer xenografts in mice as a single agent and in combination with mitoxantrone. GTI also adds to the anti-tumour efficacy of taxanes in breast cancer xenografts in mice. A Phase 1 study of a 14 day continuous infusion of GTI in patients with solid tumours showed no dose limiting toxicities at doses up to 210.9 mg/m2/day. The objective of this dose escalation phase of a phase I/II study was to define a safe phase II dose of GTI in combination with docetaxel (D) in men with metastatic hormone refractory prostate cancer (HRPC). Methods: Men with metastatic HRPC were enrolled at 3 centres in Canada. GTI was given as a 14d continuous IV infusion every 21d with D IV infusion started 2 hrs prior to the end of the GTI infusion. Planned dose escalation cohorts are summarized in table . Results: 13 men were enrolled to the 3 cohorts. All patients are evaluable for toxicity. There was one possible DLT - an episode of grade 4 neutropenia reported at cycle 2 day 1 in the highest dose cohort - but the duration of neutropenia could not be confirmed. 3 additional patients were accrued to that cohort with no DLTs. The most common gr 3/4 toxicity was attributable to D (10 pts with Gr 3/4 neutropenia). The observed incidence of Gr 3/4 neutropenia was expected since patients had weekly CBCs. Only 1 patient had febrile neutropenia. 11 pts had fatigue (4 Gr 3) related to D and /or GTI. Other GTI attributable adverse events were Gr 1/2 including transient rises in transaminases and PTT. The pharmacokinetic data which is summarized in the table will be presented in full at the meeting. Conclusions: GTI can be given safely at its highest planned dose with standard doses of D. A Phase II evaluation of the GTI + D combination is planned for men with HRPC.

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In vitro characterization and pre-clinical pharmacokinetics of CP-870,893, a human anti-CD40 agonist antibody
Authors: Bedian V. , Donovan C. , Gardner J. , Natoli E. , Paradis T. , Alpert R. , Wang H. , Shepard R. , Wentland J. , Gladue R.
Published: March 2020

2539

Background: CD40 is expressed on B-cells, monocytes, dendritic cells, other normal tissues and tumors. Previous studies showed that CD40 stimulation enhances antigen presentation, breaks tolerance, bypasses T-cell help, and induces apoptosis in CD40pos tumor cells. We report the in vitro activity and primate pharmacokinetics of a human anti-CD40 agonist antibody, CP-870,893, currently in clinical trials for the treatment of cancer. Methods: CP-870,893 was identified as a CD40 agonist antibody by screening lead molecules generated through the Abgenix Xenomouse® platform. Agonist activity was determined using upregulation of B-cell and monocytes derived dendritic cell surface markers, as well as dendritic cell IL-12 induction. BIAcore and equilibrium binding were utilized to determine affinity, and competition studies with CD40L were conducted on BIAcore. CP-870,893 was administered to cynomolgus monkeys i.v. at various doses, serum antibody levels were evaluated over time in an ELISA assay, and B-cell markers were monitored by FACS. Results: CP-870,893 (IgG2, kappa) binds CD40 with sub-nanomolar affinity, and does not block binding of CD40L. When human whole blood is incubated with CP-870,893, upregulation of key surface molecules involved in antigen presentation (MHC Class II, CD80, CD86, CD23 and ICAM-1) is observed with an EC50 of 5–50 ng/ml. Human monocytes derived dendritic cells, when stimulated with CP-870,893, upregulate activation markers (MHC Class II, CD80 and CD83) with an EC50 of 100–300 ng/ml, and secrete high levels of IL-12p40. In the presence of a second stimulus, such as LPS, human dendritic cells also secreted bioactive IL12-p70 when stimulated with CP-870,893 (EC50 ∼ 150 ng/ml). In addition, a CD40 positive human B-cell tumor line, when stimulated with CP-870,893, becomes susceptible to killing by human CTLs. In cynomolgus monkey studies, the clearance of CP-870,893 decreased with increasing dose. Circulating B-cell numbers decreased, and surface molecules were upregulated on B-cells. Conclusions: These data support the potential utility of CP-870,893 as an immune enhancing agent in cancer immunotherapy, by activating antigen presenting cells, and by enhancing the immunogenicity of CD40 positive tumor cells.

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Anti-tumor activity of partially HLA-matched irradiated blood mononuclear cells in patients with advanced malignancies
Authors: Strair R. , Gharibo M. , Taber BS K. , Kuriyan M. , Dipaola R. , Stein M. , Todd M. , Rubin A. , Lattime E. , Medina D.
Published: March 2020

2552

Background: We previously demonstrated the safety and efficacy of HLA-haploidentical blood mononuclear cells (MNCs) administered in the absence of preparative therapy to patients with metastatic renal cell carcinoma (RCC) (J Clin Oncol 21:3785–91.2003). The efficacy of this therapy is evaluated further in patients with advanced malignancies. Methods: Patients with advanced malignancies who had HLA- haploidentical donors were enrolled. Donors underwent 12–15 L leukapheresis. MNCs were processed by irradiation (25 Gy) and immediately infused. Disease evaluation was undertaken every 8 weeks. Repeat infusions were given every 8 weeks until disease progression. Results: 30 patients with diverse diagnoses, including 9 patients with a hematological malignancy were treated. Three of 13 patients with RCC had disease response, and 1 of 2 patients with melanoma had a mixed response. A patient with imatinib mesylate-refractory chronic myelogenous leukemia (CML) had a transient response. A patient with refractory acute myelogenous leukemia (AML) had disease response in conjunction with therapy. Treatment was associated with minimal toxicity and radiographic responses in patients with RCC did not occur until after 2–3 treatments. Host CD8+ cells reactive with the Pr-1 peptide of proteinase 3 were induced in association with disease response in a patient with AML. Additional studies identifying disease targets and mechanisms by which infusion of irradiated HLA-haploidentical cells are associated with disease response are ongoing. Conclusions: Disease response is seen with irradiated allogeneic MNCs administered outside the context of hematopoietic stem-cell transplantation. Evidence for anti-tumor activity is seen in isolated patients with RCC, melanoma, CML and AML. The temporal nature of the response in RCC in combination with the very limited survival of the infused cells raises the possibility of induction of a host-mediated anti-tumor effect. This hypothesis is supported by induction of host CD8+ T cells reactive with a tumor-associated peptide in a patient with AML. Given the lack of toxicity, broad availability of related haploidentical donors and the relative low financial cost, this form of cellular therapy should be developed further.

No significant financial relationships to disclose.
Activation of human peripheral blood immune cells in vitro by sodium stibogluconate (SSG) and SSG/IL-2 combination
Authors: Yi T. , Fan K. , Finke J. , Bukowski R. , Borden E. C.
Published: March 2020

2557

Background: SSG, an inhibitor of SHP-1 and SHP2 which are negative regulatory signaling tyrosine phosphatases, suppressed the growth of murine renal carcinoma (Renca) in combination with IL-2 via an interferon (IFN)-γ+ T cell-dependent mechanism (J Immunol 175:7003–8, 2005). Activation of primary human immune cells by SSG or SSG/IL-2 would provide additional supporting evidence for SSG clinical evaluation in combination with IL-2. Methods: Peripheral blood of healthy and melanoma donors were treated in vitro with SSG or SSG/IL-2 and then subjected to ELISPOT assays and FACS analysis for quantification of TH1 (IFN-γ+) and TH2 (IL-5+) cells. Results: Characterization by ELISPOT assays of peripheral blood identified IFN-γ+ cells with marked increase (9.1x) following SSG/IL-2 treatment in contrast to the modest increases induced by SSG alone (2.1x) at its clinically achievable dose (20 μg/ml) or by IL-2 (3.1x) at its Cmax of low-dose schedule (30 IU/ml). SSG at an even higher dose (100 μg/ml) was less effective alone (1.5x) or in combination with IL-2 (7.8x). IL-5+ cells were not induced by the agents individually or in combination. Induction of peripheral IFN-γ+ cells by SSG/IL-2 was detectable after 4 and 16 h. Increased effectiveness of the SSG/IL-2 combination vs. the individual agents in inducing IFN-γ+ cells in vitro was detected by ELISPOT assays in the peripheral blood from 7 healthy donors and 6 melanoma patients. FACS analysis showed that the combination induced IFN-γ+ cells in the CD4+ and CD8+ lymphocytes and also increased lymphocytes expressing the activation marker CD69 in both CD4+ (2–5x) and CD4− populations (∼ 3x). Conclusion: SSG, a compound used clinically for treatment of leishmaniasis, can interact with IL-2 in vitro to activate immune cells in peripheral blood of healthy and melanoma donors. Results provide further rationale, in addition to the mouse tumor data, for proof of concept clinical trials for effecting augmentation of IL-2 through inhibition of negative regulatory signaling phosphatases.

No significant financial relationships to disclose.
Cancer therapy with tumor cell lysate pulsed monocyte-derived dendritic cells in patients with metastastic colon cancer: Improvement by danger signals
Authors: Neβelhut T. , Lorenzen D. , Marx D. , Chang R. , Neβelhut J. , Matthes C. , Cillien N. , Stücker W. , Peters J.
Published: March 2020

2563

Background: The prognosis of colon cancers with hematogenous spread remains poor at 12–20 months. We report the use of a dendritic cell based immunotherapy to improve patient survival, especially with cell culture conditions mimicking infection. Methods: After isolating monocytes from the blood of 39 patients with metastasized colon cancer, dendritic cells were generated ex vivo in the presence of recombinant cytokines (IL-4; GM-CSF) and autologous serum. The DC loaded with tumor cell lysate were administered to the patients intradermally. Culture conditions were tested for upregulation of costimulatory molecules, downregulation of IL-10 and upregulation of IL-12 secretion by ELISPOT and fluorescence cytometry. Results: DC vaccination induced a clinical response in 9 (23%) patients with a median overall survival after onset of DC-therapy of 12 months (11–44 months for responders and 1–20 months for non-responders) and 23 months after diagnosis of metastases (16–63 months for responders and 1–23 months for non-responders). These data show that a dendritic cell based immunotherapy may prolong the patients overall survival. However, complete remissions are rare. This can be due to weak stimulation of CTL response due to insufficient antigen presentation, lack of costimulatory molecules as well as secretion of IL-10 rather than IL-12 by the DC’s. Here we could show that Toll-like receptors (TLR) ligands like Poly-I:C or lipopeptides as so called danger signals in combination with interferons can induce an upregulation of costimulatory molecules accompanied by inhibition of IL-10 and induction of IL-12 secretion in vitro. By using these culture conditions we induced a clinical complete remission of liver metastases after failure of standard therapy in one patient (overall survival after onset of DC-therapy 13 months, after diagnosis 24 months). Conclusions: According to Matzinger’s hypothesis an effective immune response occurs only by responding to a danger signal associated with infection or stress. Thus, cell culture condition should be used with TLR ligands mimicking a bacterial or viral infection. In general, a dendritic cell based immunotherapy can be successful in advanced stages of colon cancer patients.

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Identification of RGS5 as a tumor-associated antigen expressed in a broad range of human malignancies and recognized by antigen specific cytotoxic T cells
Authors: Brossart P. , Boss C. , Brauer K. , Weinschenk T. , Kanz L. , Stevanovic S. , Rammensee H. , Grünebach F.
Published: March 2020

2569

Background: Identification of tumor-associated antigens (TAA) have resulted in the development of therapeutic vaccines for the treatment of cancer. We applied an integrated functional genomics approach to identify TAA in malignant tissues of patients with renal cell carcinoma (RCC). Methods: A comparative DNA chip analysis of tumor and the corresponding non-malignant tissue from patients with RCC followed by sequencing of peptides bound to the HLA-class I molecules by mass spectrometry was applied to identify novel TAA in RCC. To confirm the immunogenicity of identified epitopes cytotoxic T lymphocytes (CTL) were generated using dendritic cells (DC). Results: RGS5 was found to be overexpressed in 16 of 29 analyzed RCC (5.7–29.9 fold) as compared to non-malignant renal tissue. Two peptides derived from RGS5 binding to either HLA-A*02 or 03 were identified. RGS5 was previously shown to be expressed in pericytes and promote tumor angiogenesis. Using RT-PCR analysis we found that RGS5 is expressed on a broad variety of tumor cells including RCC, colorectal, breast and ovarian cancer, malignant melanoma and multiple myeloma as well as in acute and chronic leukemias making this protein an interesting candidate for the development of vaccination strategies to target the tumor cells and the tumor vessels. CTL that were induced using the RGS5 peptides lysed autologous DC pulsed with the cognate peptide or transfected with in vitro transcribed RGS5 RNA as well as HLA-matched tumor cell lines. The specificity and HLA restriction was confirmed using blocking monoclonal antibodies and in cold-target inhibition assays. We next utilized DC transfected with RGS5 RNA to generate specific CTL. Using this approach we confirmed the processing and presentation of the identified peptides by malignant cells. These CTL lysed tumor cells in antigen specific manner while sparing non-malignant cells. In an ongoing vaccine trial RGS5 peptides could safely be given to RCC patients resulting in the induction of peptide specific CTL responses. Conclusions: Our results demonstrate that RGS5 is a novel tumor rejection antigen expressed in a wide range of malignancies that can be applied to target malignant cells and tumor angiogenesis.

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Optimizing targeted agents development in pancreatic cancer: A fine-needle aspirate biopsy (FNAB) based ex vivo and in vivo assay
Authors: Rubio-Viqueira B. , Mezzadra H. , Iacobuzio-Donahue C. , Jimeno A. , Zhang X. , Maitra A. , Altiok S. , Hidalgo M.
Published: March 2020

3002

Background: While targeted therapeutics effectively inhibit tumor growth with less toxicity than conventional chemotherapy, in general cancer patients show highly variable responses to them. This study aimed to develop and validate methods that permit the prediction and assessment of the efficacy of targeted therapeutics by using tumor fine needle aspiration biopsy (FNAB) samples Methods: A series of human primary pancreatic cancer were xenografted in nude mice by implanting tumor materials from surgically resected pancreatic cancer patients. Animals were treated with an epidermal growth factor receptor (EGFR) inhibitor, erlotinib, and the mTOR inhibitor temsirolimus or vehicle. Tumors were sampled by FNAB prior to (baseline) and after therapy. Tumor cells obtained by FNAB at baseline were exposed to erlotinib or temsirolimus for six hours ex vivo to prospectively predict tumor response. The degree of inhibition in phosphorylation of ERK1/2 and S6-Ribosomal Protein (S6-RP) downstream to EGFR and mTOR, respectively, were assessed by Western blot to determine the efficacy of the targeted drugs ex vivo and in vivo. The results were correlated with drug-mediated changes in tumor volumes. The effect of drugs on protein phosphorylation ex vivo was also analyzed in patient FNAB samples to show the clinical feasibility of the approach. Results: The degree of inhibition of target pathway activity in tumor cells treated with erlotinib or temsirolimus ex vivo closely correlates with tumor sensitivity in vivo. Xenograft animals sensitive to therapy showed the highest average inhibition of target protein phosphorylation in ex vivo assays, whereas tumors resistant to drugs gave the lowest one. Patient tumor samples yielded adequately cellular material to analyze the effect of erlotinib and temsirolimus in ex vivo assays. Conclusions: FNAB can be used to test ex vivo tumor response to targeted agents. These effects predicted pharmacodynamic as well as antitumor effects in a serie of pancreatic cancer xenografts. Implementation of these novel methods in clinical trials may be useful to identify patients that are likely to benefit from targeted therapeutics before being exposed to drugs and their potential side effects.

No significant financial relationships to disclose.
Enzymatically-targeted 131I therapy for herpesvirus-associated malignancies
Authors: Fu D. , Lemas V. , Foss C. , Fox J. , Pomper M. , Ambinder R. F.
Published: March 2020

3010

Background: Epstein-Barr virus (EBV) and Kaposi’s sarcoma herpesvirus (KSHV) are associated with tumors including AIDS-related lymphoma, Burkitt’s lymphoma, Hodgkin’s lymphoma, nasopharyngeal carcinoma and Kaposi’s sarcoma. Both viruses encode kinases that selectively phosphorylate nucleoside analogs such as ganciclovir and FIAU. We hypothesized that it might be possible to use the viral enzymes to specifically concentrate 131I-FIAU in tumor cells harboring virus and thus deliver therapeutic radiation. Bortezomib is a potent stimulator of viral kinase expression in gammaherpesvirus tumor cell lines. Methods: We evaluated the effect of 131I-FIAU on human cancer xenografts in SCID mice. These included a tumor line engineered to constitutively express the EBV thymidine kinase (EBVTK), and a control engineered with a sham vector (SHAM), as well 2 EBV(+) Burkitt’s lymphoma (BL) lines, and 1 KSHV(+) primary effusion lymphoma (PEL) cell line. Mice were treated with buffer, bortezomib 2 ug/gm, or I-FIAU, or I-FIAU and bortezomib in combination. For imaging, mice, 125I -FIAU and SPECT were used. For therapy, 131I-FIAU was used and tumor dimensions were monitored with calipers. Results: Treatment with buffer had no effect on on 3 EBVTK tumors and 3 SHAM tumors all of which increased in volume. Treatment with 1.4 mCi 131I-FIAU alone led to tumor responses (>90% volume reduction at 10 days) in 3/3 mice with EBVTK tumors and 0/3 mice with SHAM tumors. Treatment with 131I-FIAU alone had no effect on BL (0/3) or PEL (0/9) xenografts and all tumors increased in volume. Treatment with bortezomib induced modest responses in all tumors but had no greater effect on EBVTK tumors than SHAM tumors. However, treatment with bortezomib and 131I-FIAU led to marked tumor regression (>95%) in each of the virus-associated tumors (3/3 BL, 9/9 PEL). SPECT imaging with 125I-FIAU showed selective concentration of radiolabel in tumor tissue in the EBVTK tumor (2/2) and in viral tumors (6/6) when bortezomib was administered. There was no selective concentration in the absence of bortezomib treatment in the viral tumors (0/8). Conclusions: Treatment with bortezomib leads to selective concentration of labeled FIAU in the herpesvirus-associated tumor xenografts evaluated and to regression of tumor when the isotope is 131I.

No significant financial relationships to disclose.
A phase I dose escalation study of BIBW 2992, an irreversible dual EGFR/HER2 receptor tyrosine kinase inhibitor, in patients with advanced solid tumours
Authors: Shaw H. , Plummer R. , Vidal L. , Perrett R. , Pilkington M. , Temple G. , Fong P. , Amelsberg A. , Calvert H. , De Bono J.
Published: March 2020

3027

Background: BIBW 2992 is a novel, potent, orally bioavailable irreversible inhibitor of EGFR and HER2 receptor tyrosine kinases with IC50 values of 0.5 and 14 nM, respectively. Methods: Patients (pts) with advanced solid malignancies were enrolled. BIBW 2992 was given orally as a continuous once daily dose from 10 mg, doubled in successive cohorts until drug-related toxicity > grade 2, when escalation of no more than 50% was allowed. All pts had pharmacokinetic sampling and pre- and post-treatment skin biopsies. DNA sequencing of tumour cell EGFR and HER2 was performed on pts achieving objective response. Results: Twenty-six pts were treated (14 M/11 F). Median age: 53 (range: 30–68). ECOG PS 0/1: 7/19. 19 pts completing ≥ 28 days were evaluable for adverse events (AEs). Three dose-limiting toxicities (DLT) were seen. One pt with HER2+ breast cancer who had previously received trastuzumab and lapatinib treated at 30 mg of BIBW 2992 developed dyspnoea with radiological interstitial changes, which fully recovered on drug discontinuation. The 2 other DLTs were grade 3 acneiform skin rash, at doses of 40 mg and 50 mg daily. AEs resolved on drug discontinuation and the pts were dose reduced to 30 mg and 40 mg, respectively. The 50 mg dose level is being expanded. One pt treated at 50mg developed grade 3 diarrhoea in cycle 2 and was dose reduced to 40mg with resolution of the AE. Other AEs were mild (CTCAE v3 grade 1 or 2); nausea, diarrhoea, mucositis and fatigue. Two female pts with lung adenocarcinoma treated with 10 mg and 40 mg daily had confirmed partial responses (PR) and remain on study after 14 and 5 months, respectively. The pt with PR at 10mg has a complex heterozygous EGFR mutation in tumour cells, including a deletion and missense mutations of 4-amino acids (WT: KELREATSPKANKEILD; Patient: KEP—-SPRANKEILD) in the kinase domain, but a wildtype HER2 domain. One pt with cervical and one pt with colorectal cancer had stabilization of disease and tumour markers for 6 and 4 months, respectively. Conclusions: BIBW 2992 shows promising activity with skin rash as the main AE. Maintained PR seen in lung adenocarcinoma indicates effective mutated EGFR kinase modulation at the lowest evaluated dose level. Exploration of BIBW 2992 in Phase II is warranted.

[Table: see text]
Topical vitamin K3 (Vit K3, Menadione) prevents erlotinib and cetuximab-induced EGFR inhibition in the skin
Authors: Perez-Soler R. , Zou Y. , Li T. , Tornos C. , Ling Y.
Published: March 2020

3036

Background: The EGFR inhibitors erlotinib and cetuximab cause skin toxicity in about 75% of patients. The occurrence of skin toxicity correlates with clinical benefit. About half the patients with skin toxicity report significant discomfort. There are currently no scientifically based or proven methods for preventing or treating effectively the skin toxicity secondary to EGFR inhibitors. Methods: We screened a number of EGFR activators and phosphatase inhibitors for their ability to abrogate EGFR inhibition secondary to erlotinib and cetuximab in A431 cells. P-EGFR expression was assessed by western blot analysis. Vit K3 was selected for further in vivo studies. The skin toxicity secondary to the topical application of Vit K3 was evaluated in nude mice. The highest non-toxic concentration was used to determine the ability of topical Vit K3 to prevent EGFR inhibition in the skin of nude mice treated with EGFR inhibitors. Results: Vit K3 was the most potent EGFR activator identified, the maximum effect being observed at concentrations of 0.1–0.5 mM. In the presence of erlotinib or cetuximab, 0.1- 0.5 mM Vit K3 completely prevented EGFR inhibition. The maximum non-toxic concentration of Vit K3 applied topically BID × 10 days to nude mice was 15 mM. At this concentration, topical Vit K3 caused P-EGFR upregulation in the skin. In animals treated with erlotinib (100 mg/kg daily × 5 days), there was no detectable P-EGFR expression in the skin not treated with topical Vit K3 whereas P-EGFR expression was completely restored in the skin treated BID x 5 days with topical Vit K3. Conclusions: Vit K3 is one of the first reported agents to prevent/reverse EGFR inhibition secondary to anti-EGFR agents. The results strongly justify the development of a topical formulation of Vit K3 to treat and prevent the cutaneous toxicity secondary to EGFR inhibitors.

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Toxicity results and early outcome data on a randomized phase II study of docetaxel ± bevacizumab for locally advanced, unresectable breast cancer
Authors: Lyons J. A. , Silverman P. , Remick S. , Chen H. , Leeming R. , Shenk R. , Fu P. , Dumadag L. , Escuro K. , Overmoyer B.
Published: March 2020

3049

Background: Preclinical models of combination angiogenesis inhibitor bevacizumab (rhuMAbVEGF) and docetaxel demonstrate synergistic suppression of capillary vessel formation. Based upon these data, we developed a randomized phase II trial in order to evaluate the vascular effects on tumor regression with combination bevacizumab/docetaxel vs. docetaxel in the treatment of locally advanced breast cancer. Methods: 49 patients (pts) were randomized to receive neoadjuvant therapy with bevacizumab (10 mg/kg qowk) and docetaxel (two 8-week cycles of 35 mg/m2 weekly x 6 with a 2 wk break) (BD=24) or docetaxel (D=25) alone. Eligible pts had locally unresectable breast cancer with (n=6) or without distant metastasis (n=43); 16 patients presented with inflammatory breast cancer. Pts whose disease responded, sequentially underwent definitive surgery (4 weeks after BD or D), radiation, 4 cycles of conventional Adriamycin/cyclophosphamide, and tamoxifen or anastrazole (if ER/PR+). Results: Among the 49 pts: 7 clinical CRs, 32 PRs, 5 NR, and 5 PD. Of the 37 pts who underwent surgery: the median number of pathologically positive lymph nodes (LN) was 1 (BD=6, D=1; p=0.228); range 0–20; 43% were LN negative. Neoadjuvant treatment toxicity for both arms was acceptable with no significant differences between the two arms. Grade 4 toxicity included BD - new papillary thyroid cancer (1), neutropenia (1), hyperuricemia (1) and colon perforation (1); and D: - hyperglycemia (1) and hyperuricemia (1). 21 patients in each arm experienced a grade 3 toxicity. There were no episodes of uncontrolled hypertension, proteinuria, or thrombosis. Delayed wound healing (unable to start radiation w/in 6 weeks of surgery) occurred in 8 pts: BD=5; D=3 (p=0.691). Only 1 pt (D) experienced a change in LVEF by > 15% or below the institution’s lower limit of normal. Conclusions: Neoadjuvant therapy for locally advanced breast cancer using docetaxel with bevacizumab is well tolerated. Further studies are required to determine the added efficacy from bevacizumab. Correlative studies on impact of treatment on angiogenesis will be reported separately. (Sponsored by grants: K23CA 87725–01, M01 RR 00080, UO1 CA 62502, 5P30 CA43703-NCI/AVON, Aventis)

No significant financial relationships to disclose.
A phase I study of reolysin given intravenously to patients with advanced malignancies
Authors: Vidal L. , Pandha H. , Spicer J. , Harrington K. J. , Allen S. , Leader D. , Coffey M. , Thompson B. , Kaye S. , De-Bono J.
Published: March 2020

3064

Background: Reovirus is a double-stranded RNA virus with minimal pathogenicity in humans that selectively replicates in cells with activated Ras. Activated Ras inhibits the anti-viral effects of double stranded RNA-activated protein kinase (PKR). Reovirus serotype 3 Dearing has selective antitumor activity, in vitro and in tumour xenograft models. Methods: Reolysin was administered as a 1-hr IV infusion every 4-weeks initially for one day; then 3 days then 5-days every 4 weeks. The starting dose was 1×108 tissue culture infectious dose (TCID50) increasing in successive cohorts until observation of drug-related toxicity ≥ grade 2. Endpoints were safety, viral replication, viral shedding, evaluation of immune response and antitumor activity. Results: 24 patients (pts) (median age 60; ECOG 1, 16 males) have been entered into the first 7 cohorts at the following dose levels: 1×108 for 1-day, 1×108 for 3-days and 1×108, 3×108, 1×109, 3×109 and 1×1010 TCID50 for 5-days. A maximum tolerated dose (MTD) has not been reached and no dose-limiting toxicities have been observed. Toxicities have been mild (grade 1 or 2) and have included chills, fever, headache, runny nose, fatigue and myelosuppression. Reverse transcription polymerase chain reaction (RT-PCR) studies of blood, urine, stool and sputum post reovirus administration were negative for viral shedding for all treated pts. All but one pt had neutralising anti-reovirus antibodies detectable pre-treatment. Titres increased after 1-week of treatment and remained high during subsequent courses of treatment. Two pts with metastatic colorectal cancer treated at 3×108 and 3×109 TCID50 had CEA tumour marker reduction by 60% and 27% receiving 6 and 3 courses of treatment respectively. One pt with metastatic pancreatic cancer received 4 courses of treatment with stable disease. One pt with metastatic prostate cancer had a 50% decrease in PSA after treatment at 3×109 TCID50, with evidence of tumor necrosis on CT scanning. Intratumoral reovirus replication has been detected by electron microscopy in tumur biopsies. Conclusions: Reolysin is well tolerated with minimal toxicity. No viral shedding has been detected. Virus-induced tumor necrosis associated with intratumor viral replication after systemic delivery has been observed.

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Phase I study of intravenous CCI-779 in combination with bryostatin-1 in solid tumors
Authors: Haas N. B. , Lewis N. , Cohen R. B. , Malizzia L. , Einarson M. B. , Walker M. A. , Kim T. , Von Mehren M. , Hudes G. R.
Published: March 2020

3067

Background: mTOR regulates translation of messenger RNA critical for angiogenesis and cell growth. CCI-779 inhibits mTOR signaling through p70S6 kinase (S6K), which phosphorylates the ribosomal S6 protein (S6). CCI-779 is active in renal carcinoma (RCC) and other solid tumors. Based on our finding that bryostatin-1 (bryo) inhibits S6K, and that CCI-779 + Bryo additively inhibit S6K and RCC growth in vitro, we initiated a phase I trial to determine the maximum tolerable (MTD) doses and dose-limiting toxicities of this novel combination in patients with solid tumors. Methods: Bryo (20 μg/m2) was administered over 60 minutes IV followed by CCI-779 (10 and 15 mg, planned escalation to 75 mg) IV over 30 minutes, both weekly for 3 of 4 weeks. Serum and peripheral blood mononuclear cell (PBMC) samples were collected for analysis of pharmacokinetics and markers of mTOR inhibition (phospho-S6 and p21waf1). Results: Nine patients (median age 57, 6 RCC, 2 sarcoma and 1 neuroendocrine) are evaluable for toxicity (total cycles=27 cycles, median 4, range 1–7) at the initial 2 dose levels (Bryo 20 μg/m2, CCI-779 10 mg and 15 mg). Cycle 1 toxicity includes grade 3 hypophosphatemia and myelosuppression in one patient each and grade 2 fatigue, stomatitis, and anemia. One patient developed reversible grade 3 pneumonitis after 7 cycles. Of 7 patients with therapy-resistant disease, 5 (4 RCC and 1 sarcoma) had stable disease lasting up to 7 months. A therapy-naive patient with RCC has 23% tumor reduction after 2 cycles at dose level 2 (CCI-779=15 mg). PBMC proteins from 5 patients show consistent decreases in phospho-S6 at 2–6 hours-post treatment compared with pre-treatment baseline measurement, with recovery by 24–72 hours after dosing. In parallel with inhibition of phosphorylation of S6, PBMC levels of p21waf1 were completely inhibited in all 5 patients. Conclusions: The combination of bryo and CCI-779 is feasible, with antitumor activity in RCC and mTOR pathway inhibition observed at submaximal doses. Dose escalation is continuing.

No significant financial relationships to disclose.
Phase I study of BAY 57–9352, a VEGFR-2 inhibitor, in cycles of 14 days on/7 days off in patients with advanced solid tumors
Authors: Strumberg D. , Mross K. , Scheulen M. , Frost A. , Hedbom S. , Rajagopalan P. , Voliotis D. , Wensing G. , Christensen O.
Published: March 2020

3089

Background: BAY 57–9352 (BAY) inhibits the VEGFR-2 and VEGFR-3 tyrosine kinases, in addition to PDGFR-β and c-kit. BAY showed anti-tumor activity in colon, breast, pancreatic, and NSCLC preclinical models. Methods: This study investigated the safety, pharmacokinetics (PK), and pharmacodynamics (PD) of oral BAY in 14 days on/7 days off treatment cycles until discontinuation due to toxicity or progression. PK was assessed on days 1 and 14 of cycle 1. Dynamic contrast-enhanced MRI (DCE-MRI) was performed at baseline and in cycles 1–3. Plasma biomarkers (VEGF, sVEGFR-2) were assessed at each cycle. Results: 77 patients (pts) were enrolled at doses from 20 mg once daily to 1500 mg twice daily (bid). Common tumor types were CRC (31%) and RCC (16%). Frequent drug-related adverse events (≥5% pts) were hypertension (all grades, 11 pts [14%]; grade 3, 10 [13%]), hoarseness (10 [13%], only grade 1/2), anorexia (7 [9%], only grade 1/2), myalgia (6 [8%], only grade 1/2), fatigue (5 [6%], only grade 1/2), and diarrhea (all grades, 4 [5%]; grade 3, 1 [1%]). Hypertension in cycle 1 caused a dose reduction/interruption in 3 pts. One pt had diarrhea in cycle 7 requiring a dose reduction. One pt discontinued due to fatigue in cycle 8. BAY AUC increased dose-proportionally up to 900 mg bid. As measured by DCE-MRI, pts with CRC showed the greatest decrease in iAUC60 of Gd-DTPA, a parameter related to tumor blood flow and permeability. One pt with RCC achieved a partial response (PR); another had significant tumor shrinkage not reaching formal criteria (RECIST) for PR. PD responses (VEGF and sVEGFR-2 levels) increased in a dose-dependent manner up to 900 mg bid. Conclusions: The MTD was not reached for BAY up to 1500 mg bid in a 14 days on/7 days off schedule. BAY had a favorable safety profile. Tumor efficacy was shown in individual patients. The PD effect of BAY was demonstrated by DCE-MRI and plasma biomarkers. The recommended dose for Phase II evaluations is 900 mg bid.

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Evaluation of gefitinib biological effects in patients with solid tumors amenable to sequential biopsies—Final results
Authors: Kulesza P. , Brahmer J. R. , Jimeno A. , Baker S. D. , Spitz A. , Li J. , Rudek M. A. , Messersmith W. A. , Hidalgo M.
Published: March 2020

3090

Background: Biological effects of gefitinib are not well understood. The goals of this study are to assess the effects of gefitinib on EGFR signaling pathways in tumor and normal tissue, as well as the relationship between pharmacologic and pharmacodynamic (PD) endpoints. Methods: Patients (pts) with a diagnosis of a solid tumor and amenable to serial biopsies received gefitinib in two sequential cohorts of 12 pts each at doses of 250 or 500 mg orally once a day. Biopsies of tumor and skin as well as plasma and oral buccal swabs were collected at baseline and 28 days after treatment for correlative studies. Plasma samples for pharmacokinetic studies were obtained on d1 (from 0–8h), and pre-treatment trough samples were obtained on d2, d3, d8, d15, d22, and d28 of cycle 1. Treatment was continued unless severe drug related toxicity developed or until disease progression. Results: 27 pts (15 M/12F; median age of 61, range 29–77) were treated (13 on 250 mg and 14 on 500 mg). Three pts withdrew prior to d28 biopsy. One pt with metastatic squamous cell skin cancer had a partial response (PR). Of the 4 pts with stable disease (SD) (2-carcinoid, 1-lung, 1-renal), the median time on study was 6 mo (range 3–7 mo). Grade 1–3 toxicities included rash, diarrhea, nausea, anorexia, and fatigue. Plasma exposure to gefitinib was highly variable. At 250 mg, gefitinib average pre-treatment trough concentrations (Css,min) during 28 days of treatment varied 19-fold (mean, 406 ng/ml; range, 104 to 1846 ng/ml); at 500 mg, Css,min varied 11-fold (mean, 502 ng/ml; range, 145 to 1590 ng/ml). Quantitative immunohistochemical staining revealed a correlation between a lower baseline level of tumor Ki-67 and benefit from treatment (PR + SD) (p=0.048). While the change in tumor pMAPK staining (change from baseline) was not different between the two dose levels, Ki-67 change trended greater in the pts on the 500 mg dose (p=0.06). Change from baseline in pMAPK correlated with the development of diarrhea (p=0.03). EGFR (CA) repeat polymorphism length was not related to treatment benefit, toxicity, or changes in pMAPK or Ki-67. Conclusions: The preliminary PD findings are provocative. Further testing of tumor and normal tissues is ongoing.

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OPTIMOX2, a large randomized phase II study of maintenance therapy or chemotherapy-free intervals (CFI) after FOLFOX in patients with metastatic colorectal cancer (MRC). A GERCOR study
Authors: Maindrault-Goebel F. , Lledo G. , Chibaudel B. , Mineur L. , Andre T. , Bennamoun M. , Mabro M. , Artru P. , Louvet C. , De Gramont A.
Published: March 2020

3504

Background: The OPTIMOX1 study (JCO 2006) has shown that the strategy of 6 cycles of FOLFOX7 followed by maintenance therapy and FOLFOX reintroduction was as active and better tolerated than FOLFOX4 until progression. The aim of the OPTIMOX2 study was to evaluate a complete stop of chemotherapy after 6 bimonthly cycles of FOLFOX. Methods: Initially planned as a phase III study, OPTIMOX2 was downgraded to a large phase II study since the availability of bevacizumab. Patients (pts) were randomized between an OPTIMOX1 arm: 6 cycles of FOLFOX7 followed by LV5FU until progression and reintroduction of FOLFOX7, and the OPTIMOX2 arm: 6 cycles of FOLFOX7, complete stop of chemotherapy and reintroduction of FOLFOX7 before the tumor progression reached the baseline measures. Results: 187/200 planned pts were included between Feb 2004 and Nov 2005. Response rates were (OPTIMOX1 arm/OPTIMOX2 arm): CR 2%/2%, PR 54%/51%, stable 24%/33%, progression 11%/7%, non assessable 9%/7%. Median PFS were (OPTIMOX1 arm/OPTIMOX2 arm) 36/28 weeks (p=.01), PFS in responders 41/30 weeks (p=.001), PFS in stable patients 34/26 weeks (p=.23). Median duration of disease control (DDC), addition of PFS of first FOLFOX7 administration plus PFS of FOLFOX reintroduction if no progression at first evaluation, was 41 weeks in the OPTIMOX1 arm and 36 in the OPTIMOX2 arm, p=.17. Median duration of chemotherapy-free interval in the OPTIMOX2 arm was 25 weeks (5.7 months). Conclusions: Maintenance LV5FU therapy prolongs PFS. The quality of life of almost 6 months CFI can balance a small advantage in DDC for maintenance therapy. Our next goal is to evaluate maintenance therapy with targeted agents alone.

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