bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential expression of the 150-kd oxygen-regulated protein (ORP150) in benign, pre-malignant and malignant breast tissue
Authors: Stojadinovic A. , Hooke J. , Shriver C. D. , Kovatich A. , Nissan A. , Ponniah S. , Peoples G. E. , Moroni M.
Published: March 2020

10770

Background: Heat shock protein, ORP150, plays a role in hypoxia/ischemia and angiogenesis. Preliminary studies demonstrate increased ORP 150 expression in human cancer, and associate its over expression with aggressive tumor biology. This study further evaluates ORP150 expression in different stages of breast cancer, such as benign, pre-malignant and malignant breast lesions and correlates it with clinical-pathological data. Methods: Sixty-six prospectively collected paraffin-embedded breast tissue sections were reviewed for diagnostic confirmation (normal, n=25; DCIS, n=20; invasive breast cancer (Ca), n=21) and stained using ORP150 antibody immunohistochemistry (IHC). Antibody-staining levels in the benign, DCIS and invasive component of each lesion were reviewed independently by two pathologists and scored: 0 (lowest) to 3+ (highest). Clinical-pathological data was compared with ORP150 staining using ANOVA or T-Test as appropriate with significance determined by p<0.05. Results: Significant differential ORP150 staining was detected in benign-normal versus benign adjacent to invasive cancer, as well as in benign adjacent to DCIS versus benign adjacent to invasive cancer. ORP150 expression in the invasive portion of the breast cancer correlated significantly with tumor grade and absence of hormone receptor expression, presence of lymphovascular invasion and lymph node metastasis. Conclusions: ORP150 expression in breast cancer is associated with poor prognostic histological factors. As ORP150 is differentially expressed in benign tissue adjacent to invasive cancer, further study is warranted to determine its utility in detecting occult invasive cancer within benign biopsy specimens, as well as its putative role in tumor-stromal cell interactions.

[Table: see text]

No significant financial relationships to disclose.
An IMRT technique to increase therapeutic ratio of breast irradiation in patients with early stage left breast cancer: Limiting second malignancies
Authors: Ahmed R. S. , Fiveash J. B. , Popple R. A. , Spencer S. A. , De Los Santos J. F.
Published: March 2020

10791

Background: The clinical application of IMRT for adjuvant treatment of breast cancer has been the subject of increasing study in recent years. IMRT plans have improved target coverage and reduced dose inhomogeneities observed within the breast in standard plans. IMRT was able to reduce doses delivered to the heart, lungs, and right breast at clinically significant doses, but this has been at the cost of larger volumes of low dose radiation to these structures and thus, increasing the risk for second malignancy. Our goal was to develop an IMRT beam arrangement that did not result in additional low dose spill to risk organs while maintaining equal or better target coverage. Methods: Five patients with early stage left-sided breast cancer, who underwent breast conservation surgery, and adjuvant radiation using standard wedged tangential fields, were chosen for this comparative study. An IMRT plan consisting of 6 tangential beams (3 medial and 3 lateral) was generated by using the gantry, collimator and table angles of the standard plan used for the conventional radiation (CRT), and moving the table +10 and −10 degrees on each side. The prescription dose for both CRT and IMRT plans was 45 Gy, 1.8 Gy/fraction, prescribed to the isocenter which was placed near the center of the breast. Results: IMRT plans provided significantly better coverage of the left breast than CRT plans, (p=0.03). Although dose heterogeneity was greater with the IMRT plans, the difference was not significant (p = 0.68). The mean volumes of the heart, lung, and right breast were lower in patients planned with IMRT at all dose levels from 5% to 100% dose (5% increments). This difference was significant for volumes receiving 2.25 Gy for the heart (p = 0.003), volumes receiving 2.25 Gy, 4.5 Gy, 6.75 Gy, 33.75 Gy, 36 Gy, 38.25 Gy, and 42.75 Gy for the lung (p = 0.014, 0.04, 0.044, 0.05, 0.049, 0.045, 0.05, respectively). Surprisingly, breast IMRT resulted in significantly lower right breast volumes irradiated at all dose levels compared to CRT. Conclusions: A six-tangential field IMRT technique achieved significantly better left breast coverage while maintaining lower doses to risk organs at all dose levels and therefore, reducing the potential for induction of a second malignancy.

No significant financial relationships to disclose.
Acute toxicity analysis of patients receiving surgery, Gliadel wafer implantation, and postoperative daily temozolomide with radiation therapy for primary high-grade glioma
Authors: Heery C. R. , Desjardins A. , Quinn J. A. , Rich J. N. , Gururangan S. , Vredenburgh J. J. , Friedman A. H. , Reardon D. A. , Friedman H. S.
Published: March 2020

11504

Background: Treatment of patients with newly diagnosed malignant glioma using Gliadel wafer implantation at initial surgery has been shown to increase survival (Westphal et al 2003). Similarly, administration of temozolomide during and after radiotherapy has also been shown to increase survival in this patient population (Stupp et al 2005). Accordingly use of both Gliadel and temozolomide may be advantageous for these patients although it is possible that the toxicity of these two approaches used together might be prohibitive. Methods: The Preston Robert Tisch Brain Tumor Center at Duke has occasionally treated with this approach over the last several years, and we now present an analysis of the observed acute toxicity. We retrospectively reviewed the Duke patients treated with surgery plus Gliadel wafer placement followed by daily temozolomide (75 mg/m2-150 mg/m2) and radiation therapy. Results: Of 28 patients reviewed, four patients were diagnosed with AA (WHO grade III), two patients were diagnosed with AO (WHO grade III) and the remaining 22 patients were diagnosed with glioblastoma multiforme (WHO grade IV). Two of the 28 7.1%) patients experienced grade 3 or 4 hematologic toxicity during radiation and daily temozolomide therapy. This is similar to the 7% of patients found to have hematologic toxicity reported by Stupp et al (2005). Three patients (10.7%) had grade 3 or 4 seizure activity. Two patients (7.1%) had grade 4 pulmonary emboli. No events of cerebral edema or wound complications were noted in this review of patient events following Gliadel wafer placement. Conclusions: In summary, the addition of Gliadel wafer placement at the time of surgery followed by radiation therapy with concurrent daily low dose temozolomide does not appear to have significant acute toxicity over that observed with radiation therapy and daily temozolomide. Future formal trials combining these therapeutic strategies may allow evaluation of the possible survival advantage associated with this approach.

[Table: see text]
Treatment with temozolomide for malignant gliomas: Is rechallenge with alternative dosing regimens successful?
Authors: Balmaceda C. , Pannullo S. , Serventi J. , Sisti M. , Bruce J. , McKhann G. , Lai R. , Rosenfeld S. , Fine R.
Published: March 2020

11514

Background: Temozolomide (TMZ) is an alkylating agent with activity against malignant gliomas. A variety of dosing schedules has been used including: 5 days on/21 days off (200 mg/m2/d), bid dosing (initial bolus of 200 mg/m2 followed by bid dosing of 90 mg/m2 × 9 doses), and 7 days on/7 days off (150 mg/m2/d). It is not known which regimens are the most effective. Furthermore, it is not known whether patients failing one schedule will respond to alternative ones. Materials and Methods: We report on a retrospective series of 8 patients (7 M, 1 F), who were treated with TMZ at least twice. Mean age at recurrence was 48 (26–58). Pathology revealed GBM (3), AA (4), and AO (1). 7 patients had received prior XRT. 7 patients had a local (L) recurrence at the time of retreatment with TMZ, and one patient had leptomeningeal (LM) and L recurrence. 7 pts received TMZ alone both at the time of the first recurrence and at the time of rechallenge. One patient had received TMZ concommitant with XRT and TMZ alone at the time of rechallenge. Results: See Table . Toxicity was mild and not different than that seen in patients treated with the first course of TMZ. Conclusions: While the number of patients is limited, some observations can be made: 1) patients can respond to TMZ at rechallenge, particularly if a prior response to TMZ had been observed. 2) Some patients who fail or respond modestly to one regimen may achieve a better response to alternative dosing schedules. Further studies need to address whether one regimen of TMZ given at rechallenge allows an improved survival as compared with other regimens without sacrificing safety.

[Table: see text]

[Table: see text]
Combination of bevacizumab, a monoclonal antibody to vascular endothelial growth factor (VEGF), and temozolomide: Study of cases
Authors: Kirkpatrick J. , Desjardins A. , Vredenburgh J. J. , Quinn J. A. , Rich J. N. , Sathornsumetee S. , Gururangan S. , Friedman A. H. , Friedman H. S. , Reardon D. A.
Published: March 2020

11522

Background: The prognosis for glioblastoma multiforme remains poor. Survival is generally limited to less than 1 year. Currently available standard treatments have not allowed, thus far, to prolong survival significantly. Response rates observed in clinical trials evaluating glioblastoma multiforme are usually less than 20%. Knowing that malignant gliomas have high concentrations of VEGF receptors, and the higher the VEGF receptor concentration, the worse the prognosis, we decided to evaluate the efficacy of bevacizumab in malignant brain tumor patients. Bevacizumab is a humanized IgG1 monoclonal antibody to VEGF, which is synergistic with chemotherapy for most malignancies. We performed a phase II study combining bevacizumab with irinotecan for patient with malignant gliomas and observed an unprecedented response rate of 63%. Methods: Building of those results, we decided to treat a number of our patients with voluminous unresectable disease with bevacizumab and temozolomide as an upfront regimen. Temozolomide is an oral methylating agent known effective for primary malignant brain tumor patients. A phase III trial, first presented at the ASCO meeting of 2003, demonstrated the efficacy of temozolomide for newly diagnosed glioblastoma multiforme patients, establishing temozolomide as the new standard of care. Given the known results with temozolomide as monotherapy and the combination of bevacizumab with irinotecan, we treated patients with temozolomide and bevacizumab upfront. Results: With this new combination, some patients demonstrated dramatic improvement clinically and radiographically. The combination has been well tolerated thus far, with no incidence of hemorrhage or arterial thrombosis observed. Conclusions: Results will be updated at the time of presentation.

[Table: see text]
Phase I pharmacokinetic(PK)/food effect and safety study of satraplatin in patients (pts) with advanced solid tumors
Authors: George B. J. , Ricart A. D. , Calvo E. , Chu Q. , Sarantopoulos J. , Greene D. , Nathan F. E. , Petrone M. E. , Tolcher A. W. , Papadopoulos K. P.
Published: March 2020

12014

Background: Satraplatin (S) is a novel oral platinum analog that has demonstrated preliminary activity in hormone refractory prostate cancer (HRPC). A large (950 pts) worldwide, randomized phase III trial evaluating S as 2nd line therapy for HRPC recently completed enrollment. The current study was designed to determine the PKs of S under fasted and fed conditions, as well as treatment efficacy and toxicity. Methods: S was administered orally at 80 mg/m2/day (d) on d1–5 q 35 days, with prophylactic antiemetics. For the 1st cycle, pts were randomized to receive d1 and d5 doses of S in either the fed or fasted state when PK sampling was performed. The fed/fasted state was reversed for the 2nd cycle. Subsequent doses of S were given in the fasted state. Results: 17 pts were enrolled (9M/8F), median age 62 (range 33–78) and tumor types: HRPC (7), breast (3), 1 each with anal, parotid, leiomyosarcoma, Merkel cell, ovarian, mesothelioma, and neuroendocrine tumor. The median no. of prior regimens was 3 (range 1–8). A total of 58 cycles of S were given. Grade 3–4 hematologic toxicities (1st 2 cycles) included: neutropenia (11%), thrombocytopenia (24%) and anemia (24%). The hematologic nadir occurred during week 4. There was no grade 3–4 nausea, vomiting or diarrhea. No significant cardiac, renal, hepatic, or neurologic toxicity was observed. The peak plasma concentration (Cmax) for platinum in plasma ultrafiltrate (PUF) was decreased by 20% when S was administered following a high fat meal compared to the fasting condition. There was no effect of food on AUC0–24 hr. The time of Cmax (Tmax) was delayed in the fed state. Table shows PK parameters. One PR (HRPC) and 4 SD ≥ 5 months (breast, ovarian, parotid and HRPC) were confirmed. Conclusions: There is an effect of food on the Cmax, however the clinical implications are unknown at this time. For this reason, it is recommended that S be administered to pts in the fasting state. At the dose of 80 mg/m2/day, S was well tolerated in this group of heavily pretreated patients.

[Table: see text]

[Table: see text]
Fixed dose rate (FDR) gemcitabine (G) and capecitabine (C) in patients with advanced and/or metastatic cancer: phase I trial
Authors: Santini D. , Vincenzi B. , Virzì V. , La Cesa A. , Schiavon G. , Leoni V. , Gasparro S. , Grilli C. , Calvieri A. , Tonini G.
Published: March 2020

12016

Background: Combination of C and G has been demonstrated to be well tolerated, with apparent efficacy in patients with advanced cancers. To determine the toxicity of C plus Fixed Dose Rate (FDR) G in metastatic cancer patients, a phase I trial was conducted. Methods: This is an open-label, single-center, dose-escalating phase I study. C was administered orally according to the standard 21-day schedule: bid in equal doses (650 mg/m2 bid) for 14 days every 21 days. G was administered at a FDR of 10 mg/m2 per min in escalating durations of infusion on days 1 and 8 every 21 days. The doses of G explored were 600, 700, 800 mg/m2 infused at FDR in 60, 70 and 80 minutes, respectively. 15 pts (10 female, 5 male), aged 37–70 yr (median 66) with a variety of advanced solid tumors have been treated (11 peri-ampullary cancers, 3 colorectal cancers and 1 ovarian cancer). The FDR G dose was escalated when 3 patients in a cohort had completed two cycles of treatment without experiencing dose-limiting toxicities (DLT). The MTD was defined as the dose level at which no more than one of nine patients experienced a DLT. Results: No DLT occurred at doses of 600 and 700 mg/m2 in any of the 3 patients included at each level. At 800 mg/m2, 1 of 9 patients experienced DLT (neutropenia grade 4 with fever). The non-haematological toxicities have been generally mild or moderate in all patients (grade 2 stomatitis: 4 patients; grade 2 fatigue: 3 patients; grade 2 nausea/vomiting: 2 patients). 2 patients showed one episode of grade 4 neutropenia spontaneously regressed in 3 days. 2 patients showed one episode of grade 2 thrombocytopenia. The recommended dose for further studies is C bid in equal doses (650 mg/m2 bid) for 14 days (28 doses) plus FDR G at 800 mg/m2 infused in 80 minutes on days 1 and 8 every 21 days. Other patients with periampullary carcinoma are being evaluated at the same dose level, with an ongoing evaluation of cumulative (all cycles) toxicity and efficacy. In terms of response, we have observed 2 PR and 3 SD in the 6 pancreatic cancer patients evaluated for response after the first 3 cycles. Conclusions: C plus FDR G combination seems to be a feasible and safety approach which has demonstrated promising clinical activity.

No significant financial relationships to disclose.
Phase I study of pegylated liposomal doxorubicin (PLD) and irinotecan (I) in patients with solid tumors
Authors: Pillot G. A. , Wildi J. , Tan B. , Suresh R. , Fracasso P. , Govindan R.
Published: March 2020

12023

Background: Irinotecan (I) and doxorubicin are cytotoxic agents that interact with topoisomerase I and II respectively, and display some evidence of preclinical synergy. Pegylated liposomal doxorubicin (PLD) is a novel formulation of doxorubicin that may be more tolerable than doxorubicin. We sought to define the maximum tolerated dose (MTD) of PLD and I when given in combination. Methods: Patients (pts) with good performance status (PS 0–2) and adequate cardiac, renal, liver, and bone marrow function who had received any number of prior therapies for advanced solid tumors were eligible. Pts in the first cohort received PLD at a dose of 20 mg/m2 IV on day 1 and I 100 mg/m2 on days 1 and 8 of a 21 day cycle to a maximum of 6 cycles. Growth factor support was not permitted. Results: Eleven pts were enrolled, 6 male, 5 female. Median age of the pts was 63 years (range 19–73). There were 2 cases each of non-small cell lung cancer, renal cell carcinoma, and biliary carcinoma, and one case each of breast, pancreatic, ovarian, colon, and bladder carcinoma. Three pts were initially enrolled without dose-limiting toxicity (DLT) in cohort 1, and six pts were subsequently enrolled in cohort 2 (20 mg/m2 PLD and 125 mg/m2 I), with 2 patients experiencing DLT (neutropenia, diarrhea, and fatigue). Two additional pts have been enrolled in cohort 1 (5 pts total) without DLT, and the 6th patient will be enrolled soon to define the MTD. Grade 3 neutropenia was noted in 1 pt and grade 4 neutropenia in 2 pts. Other non-hematologic toxicities across all cycles have included grade 3 anorexia, diarrhea, nausea, and fatigue in 2 pts each, and 1 pt each with grade 3 dehydration, emesis, pneumonitis, and DVT. No cardiac toxicity has been observed. Of 6 evaluable pts, 4 experienced stable disease (1 each of NSCLC, breast, pancreas, and renal cell carcinoma). Conclusions: The combination of pegylated liposomal doxorubicin and irinotecan is feasible and well- tolerated. The maximal tolerated dose is likely to be PLD 20 mg/m2 d1 and I 100 mg/m2 d1 and 8 IV q 21 days (pending enrollment of the last patient). This study was supported by Tibotec Therapeutics, Division of Ortho Biotech.

[Table: see text]
Isolation of large numbers of human mesenchymal stem cells from discarded filters after bone marrow harvests: Implications for therapy
Authors: Mageed A. S. , Pietryga D. , Gagin S. , West R.
Published: March 2020

12501

Background: Bone Marrow (BM) is an excellent source of Mesenchymal Stem Cells (MSCs) which can be expanded and used clinically. There is a growing evidence that MSCs can promote engraftment and reduce Graft versus Host Disease (GVHD). Bone marrow harvests are passed through 850μ, 500μ and 200μ filters to trap bony spicules and clots before infusion into patients. We hypothesize that the filters would also trap a significant number of MSCs which can be useful clinically after isolation and possible expansion. Methods: Bone marrow collection kits were obtained after informed consent of the donor (n = 11). Specimens were obtained in the course of scheduled bone marrow harvests. Cells were isolated and cultured from each portion of the device. Results: An average of 1×109 total mononuclear cells was obtained from the discarded marrow collection devices. Cells isolated in this experiment conform to published data regarding MSCs. The immunophenotype was CD44+, CD73+, CD90+, CD105+, CD14−, CD34−, CD45−, HLA-DR−, negative for leukocyte lineage specific antigens and positive for Prolyl-4-Hydroxylase. The cells were morphologically consistent with published data, being bilaterally symmetrical and forming conspicuous fibroblastoid colonies which readily grew to confluence and followed differentiation pathways to both adipose osteoid tissue. Fresh cells from the kits may also contain a precursor for MSCs that is CD90− and CD73−. Sorting for this precursor population yielded colony forming units consistent with MSCs. Conclusion: This represents a novel strategy for obtaining “healthy” material for potential therapy and study.

No significant financial relationships to disclose.
Mimotopes of the disialoganglioside GD2 elicit anti-GD2 antibodies recognising GD2 on melanoma cells
Authors: Brämswig K. H. , Riemer A. B. , Förster-Waldl E. , Pollak A. , Zielinski C. C. , Pehamberger H. , Lode H. N. , Scheiner O. , Jensen-Jarolim E.
Published: March 2020

12505

Background: The disialoganglioside GD2, a carbohydrate antigen, is expressed on all tumors of neuroectodermal origin, including melanoma, sarcoma, neuroblastoma and small cell lung cancer. Due to its specific expression on tumor surfaces, GD2 is an attractive target for immunotherapy. The mouse/human chimeric antibody form ch14.18 was already applied in melanoma and neuroblastoma trials as passive immunotherapy. We aimed to replace the poorly immunogenic ganglioside with highly immunogenic peptides, in order to establish an active immunotherapy. Methods: We used the ch14.18 antibody to select GD2 mimotopes. In the present study, two mimics of the ch14.18 epitope were chosen for immunogenicity evaluation. The mimics were coupled to KLH (keyhole limpet hemocyanin) in order make them more immunogenic. Three groups of BALB/c mice were immunized i.p with the mimotope conjugates (GRL-KLH or DGG-KLH), or the carrier protein KLH alone. Results: BALB/c mice immunized with the mimotope conjugates indeed showed a specific humoral immune response towards the purified original antigen GD2 in ELISA, and also against the natural GD2 melanoma cell lysate in Western Blots. As the elicited antibodies were of the IgG isotype, the mimotope conjugates are capable of recruiting T cell help and inducing memory phenomena. Conclusion: We are able to show that an epitope of the carbohydrate antigen GD2 can successfully be translated into immunogenic peptide epitope mimics. Moreover, immunizations with these mimics induced IgG antibodies again recognizing the original antigen. We thus provide evidence that GD2 mimotopes are suitable candidates for active immunotherapy of GD2 expressing tumors. The work was supported by BioLife Science GmbH, Vienna, Austria; by project grant #10965 of the Austrian National Bank Science Fund; and by the Center of Excellence in Clinical and Experimental Oncology (CLEXO), Austrian Federal Ministry of Education, Science and Culture (GZ200.062/2-VI/1/2002). A.B. Riemer and K.H. Brämswig are recipients of Hans & Blanca Moser Fund scholarships. The work was also supported by DFG (Lo635–2) and Fördergesellschaft Kinderkrebs-Neuroblastomforschung to H.N.Lode.

[Table: see text]
Yttrium-90 (90Y) ibritumomab tiuxetan for treatment of relapsed or refractory non-hodgkin’s lymphoma: The Western Pennsylvania Cancer Institute experience
Authors: Jasthy S. , Sudan N. , Haq B. , Lister J.
Published: March 2020

12511

Background: Yttrium-90 (90Y) ibritumomab tiuxetan (Zevalin) radioimmunotherapy is an effective agent for treatment of CD20+ B-cell lymphoma (NHL).It is approved for treatment of relapsed or refractory, follicular or transformed B cell lymphoma. An overall response rate ranging from 74% to 82% has been reported. Median response duration of 11.5 to 28.1 months has been reported. Methods: We identified, retrospectively, 37 patients treated at our institution with relapsed or refractory NHL who were treated with 90Y ibritumomab tiuxetan at a dose of 0.3mCi/kg or 0.4mCi/kg. Response rate and survival were assessed. Median age was 60 years (range 28–82 years) of which 65% were male. Follicular lymphoma was present in 57% (n = 21) and diffuse large B-cell lymphoma in 22% (n = 8) of patients. Advanced disease (stage III or IV) was present in 84% (n = 31) of patients. All of the patients had received at least one prior regimen and 68% (n = 25) had received ≥ 3 regimens before 90Y ibritumomab tiuxetan. Results: Overall response rate was 65% (24/37); CR 43% (16/37); PR 19% (7/37); SD 3% (1/37). Duration of response was 13.3 months (range 4 to 48 months) with response duration greater than 12 months in 35% (n = 13) of the patients. Of responders, 67% (16/23) had failed to respond to prior immediate therapy. The only toxic event was pancytopenia, which predictably occurred after treatment. No patients died of toxicity and all toxicity was reversible. Conclusions: 90Y ibritumomab tiuxetan was well tolerated and effective for our patients.We document durable response in our patients with relapsed or refractory NHL and with advanced stage disease. Further study of ibritumomab tiuxetan using different doses and in combination with chemotherapy is warranted.

No significant financial relationships to disclose.
Immunotherapy, phase I study of immunotherapy with carcinoembryonic antigen peptide -pulsed, autologous human cultured dendritic cells in patients with advanced non-small cell lung cancer
Authors: Han B. , Huan Z. , Xiaohong F. , Rong L. , Guangli F. , Luoluo J. , Tianqing C. , Wei Z. , Jianzhong S.
Published: March 2020

12517

Background: To study toxicity tolerance and dose-effect relationship of carcinoembryuonic antigen peptide-pulsed dendritic cells in patients with advanced lung cancer. Methods: cells preparations enriched for autologous DCs were generated from the patients plastic adherent peripheral blood mononuclear cells in media supplemented with granulocyte macrophage colony -stimulating factor and interleukin-4. 37C0, 5%CO2 culture for 7–10. The DCs were loaded with carcinoembryonic antigen(CEA) in day 7, Groups of three to six patients received four weekly or biweekly i.v. infusions of the CAP-1-loaded DC in escalating dose levels of 1 × 106, 1 × 107, and 1 × 108 cells/dose. Patients with lung cancer received iv injections DCs. There were no toxicities directly referable to the treatments. Results: Total 22 patients with lung cancer received DCs immunotherapy. DCs infusion were 2.5×106-9.6×107,means 15.03×106. The early clinical trials suggest that vaccination with CEA vaccines is safe, producing few side-effects, and can lead to CEA-specific immunity. Conclusions: We conclude that it is feasible and safe to generate and administer large numbers of DCs loaded with CEA peptide to patients with lung cancer.

No significant financial relationships to disclose.
Src kinase activity in early breast cancer (EBC): A quantitative luminometric and immunohistochemical (IHC) approach
Authors: Young O. , Jones R. J. , Clack G. , Jacobs V. , Marshall A. , Fennell M. , Green T. , Dixon J. M.
Published: March 2020

13010

Background: Src kinase expression and activity is increased in many solid tumors including breast cancer, and Src kinase has a role in the regulation of cancer cell invasion. Thus, Src kinase is a potential therapeutic target. Targeting cancer cell invasion may be of particular value in early disease, although drug effects are unlikely to be measurable by conventional endpoints, for example tumor regression. This prospective study aimed to establish robust measures of Src activity in invasion-related pathways in tissue derived from EBC and to assess the variability between these measures, in order to establish a methodology for pharmacodynamic assessment of Src inhibition in clinical disease. Methods: Duplicate sets of core biopsies were taken from 30 patients, both at diagnosis and surgery. Biopsies were fixed immediately in either liquid nitrogen (luminometric analysis) or pre-cooled isopentane (IHC analysis). Formalin-fixed, paraffin-embedded tissue (FFPET) was also collected at the time of diagnosis. Levels of phospho-paxillin, phospho-focal adhesion kinase (FAK), total phosphotyrosine and total paxillin were measured in lysed tumor tissue using the Luminex system. Expression of activated Src, phospho-paxillin, phospho-FAK, total paxillin and total FAK was determined in sections of frozen tissue and FFPET, and assessed semi-quantitatively by visual examination. An analysis of variance (ANOVA) model was fitted to the Luminex data ( Table ). Results: All tumors expressed active Src, and there was a high level of expression, but low variability, of paxillin, FAK, phospho-paxillin and phospho-FAK. The subcellular localization of these markers could also be assessed in FFPET sections. Conclusion: The results indicate the feasibility of assessing the effect of a Src inhibitor on invasion-related endpoints in women awaiting surgery for EBC.

[Table: see text]

[Table: see text]
Systemic telomerase gene therapy delivered via targeted lipidic nanoparticles
Authors: Goldkorn A. , Hann B. , Hayes M. , Drummond D. C. , Kirpotin D. B. , Peth K. , Marks J. D. , Blackburn E. H. , Park J. W.
Published: March 2020

13025

Background: Most human cancers possess elevated telomerase activity that promotes proliferation. We developed 2 telomerase-targeting gene constructs: telomerase RNA containing a mutated template sequence (MT-hTer), and short interfering RNA (siRNA) targeting endogenous wild type telomerase RNA. When co-expressed in cancer cell lines and xenografts, MT-hTer/siRNA synergizes to cause rapid cell death (Li, S et al. Cancer Research 2004). Here, we tested the in vivo efficacy of MT-hTer/siRNA delivered systemically via antibody-targeted lipidic nanoparticles in a breast cancer metastasis model. Methods: Nude mice received intracardiac injection of luc+MCF7-C18 breast carcinoma cells ectopically overexpressing HER2 and luciferase. After 3 days, the mice were treated with a single 100 μg i.v. injection of either MT-hTer/siRNA-GFP or control (GFP-only) plasmid formulated into lipid-DNA nanoparticles conjugated to an internalizing anti-HER2 single chain antibody (Genosphere, TM) (Hayes, M et al. Gene Therapy 2005). A third group received saline (PBS) as a vehicle control. Cancer progression was monitored non-invasively via biophotonic imaging (Xenogen) of tumor luciferase signal. Results: By week 2 the PBS control group showed luciferase signal elevation that increased exponentially during the study. Similarly, the GFP-only control group developed significant luciferase signal elevation by week 3, with continued rise at all subsequent time points. In contrast, the MT-hTer/siRNA-GFP group experienced a decreased luciferase signal by week 2, which remained consistently low throughout the experimental time course. There were no observed treatment-related toxicities. Conclusions: In pilot in vivo studies, telomerase therapy dramatically inhibited tumor formation in mice after systemic delivery with cancer-targeting lipid-DNA nanoparticles. Expanded preclinical studies are underway.

[Table: see text]
Inhibition of histone deacetylase (HDAC) 6 sensitizes human leukemia and breast cancer cells to antagonists of heat shock protein (hsp) 90 and/or bortezomib (BZ)
Authors: Bhalla K. N. , Bali P. , Balasis M. , Fiskus W. , Pranpat M. , Rocha K. , Rao R. M. , Kumaraswamy S. , Atadja P.
Published: March 2020

13039

Background: HDAC6 is a cytosolic α-tubulin deacetylase, which shuttles misfolded polyubiquitylated proteins into aggresomes. Our studies demonstrated that the hydroxamic acid (HA) analogue pan-HDAC inhibitors (HDIs), e.g., LAQ824 and LBH589, induce α-tubulin and hsp90 acetylation, thereby inhibiting ATP-binding and chaperone function of hsp90. This promotes the polyubiquitylation and degradation of the pro-growth and pro-survival hsp90 client proteins, e.g., Bcr-Abl, mutant FLT-3, c-Raf and AKT in leukemia and Her-2 in breast cancer cells. Methods: Human CML K562 and breast cancer SKBR-3 and BT-474 cells were exposed to LAQ824 (100 nM) or LBH589 (20 to 50 nM) and/or 17-AAG (1 to 5 μM) and/or BZ (5 to 100 nM). K562 and BT-574 cells were also transfected with siRNA to HDAC6. Immunoprecipitation and/or immunoblot analysis were performed to determine the expression or binding of HDAC6 with hsp90, and of hsp90 acetylation and binding with its co-chaperones and client proteins. Immunofluorescent microscopy was utilized to estimate aggresome formation. Cellular cytotoxicity was determined by % inhibition of colony growth. Results: HDAC6 knockdown by its siRNA induced the acetylation of hsp90 and α-tubulin but not of the co-chaperones p23 and hsp40. HDAC6 binds to hsp90. Hsp90 acetylation increased its binding to biotinylated-geldanamycin (GM) and its analogue 17-AAG. Depletion of HDAC6 levels (∼70%) or its activity by treatment with HA-HDI also inhibited the binding of hsp90 to ATP, p23 and hsp90 client proteins, which was augmented by co-treatment with 17-AAG. Knockdown of HDAC6 sensitized the cells to loss of clonogenic survival induced by HA-HDI and/or BZ and/or 17-AAG. Depletion of HDAC6 led to attenuation of Her-2 levels and loss of survival of Her-2 amplified SKBR-3 and BT-474 cells. Co-localization of HDAC6 and polyubiquitylated proteins into aggresomes was enhanced by co-treatment with BZ and/or 17-AAG. This was inhibited by depletion of HDAC6 activity. Conclusions: These findings support the in vivo testing of HDAC6 inhibitors, which through inhibition of hsp90 and aggresome formation sensitizes cancer cells to 17-AAG and/or BZ induced cellular stress due to misfolded and polyubiquitylated proteins.

[Table: see text]
Preclinical development of a novel inhibitor of oncogenic B-Raf
Authors: Nolop K. , Jones W. B. , Tsai J. , Cho H. , Bremer R. , Ibrahim P. , Artis D. R. , Hirth P. , Bollag G.
Published: March 2020

13056

Background: The discovery of oncogenic B-Raf mutations in a majority of patients with metastatic melanoma, and in many tumors from patients with colorectal cancer and other cancers, presents the opportunity to develop oncogene-selective inhibitors with a favorable safety profile. Methods: Guided by co-crystallography, a novel chemical scaffold has been developed into a series of potent inhibitors of oncogenic B-Raf with selectivity versus wild-type B-Raf. From this series, a potent inhibitor has been chosen as a candidate for development. Results: Consistent with the structure-guided approach, this compound shows pronounced selectivity versus a wide array of over 70 other kinases covering all branches of the kinome. This selectivity translates to a wide cellular therapeutic index: inhibition of proliferation of a panel of cell lines bearing the V600E oncogenic B-Raf mutation occurs at IC50s ranging from 40–400 nM while inhibition of cell lines lacking oncogenic B-Raf occurs at IC50s greater than 6500 nM. This cell-based selectivity for the oncogenic B-Raf is greater than the selectivity shown in biochemical assays, supporting that tumor cells bearing the oncogenic B-Raf protein are more dependent on the MAP kinase pathway. Combination experiments with a series of cytotoxic and targeted clinical anti-cancer agents reveal multiple examples of synergistic inhibition of proliferation in vitro, and this synergy generally appears selectively in oncogenic B-Raf-bearing cells. The good oral bioavailability (F > 70%) allows for prolonged exposure in both rodents and non-rodents. Robust efficacy is evident in a murine COLO205 tumor xenograft model, with once-daily oral dosing at 20 mg/kg over 14 days resulting in > 80% inhibition of tumor growth with no effect on body weight. Conclusions: Since this compound is highly selective and targets a B-Raf variant that is absent in all non-transformed cells, it may have a broad therapeutic index, alone or in combination, for the treatment of patients with V600E oncogenic B-Raf tumors.

[Table: see text]
A phase 1 study of weekly, divided dose perifosine in patients (pts) with non-small cell lung cancer (NSCLC)
Authors: Henderson I. C. , Spigel D. R. , Nemunaitis J. J. , Richards D. A. , Liebmann J. E. , Purcell W. T. , Birch R. , Gardner L. R. , Burris III H. A.
Published: March 2020

13063

Background: Perifosine (P) is known to modulate signal transduction pathways, including Akt which is frequently constitutively activated in NSCLC. The half life of P is ∼100 hours and GI toxicities are dose limiting. Studies of a single dose weekly regimen found that the maximum tolerated dose (MTD) was 800 mg. This study was undertaken to see if a higher MTD could be reached using divided doses in NSCLC patients. Methods: Twenty NSCLC patients (pts) were given P, 300 mg, at intervals of 4–6 hours with no more than 4 doses in a 24 hour period. The starting dose was 900 mg and this was escalated by 300 mg/week in subsequent pts to an MTD of 1800 mg. Results: The median age of pts was 62 (range 39–79); 9 were male and median ECOG performance status was 1 (range 0–1). Nineteen pts had received prior chemotherapy (median 2 regimens) and 18 prior radiotherapy. Three pts were entered at each dose and the cohort expanded to 6 pts if 1 or more experienced a grade 3/4 non-hematologic toxicity (DLT) during the first week of therapy. A dose level was toxic if 2 or more pts experienced a DLT. The median time on study was 8 weeks (range 2–28). Seventy-seven percent of pts had no dose reduction (85% of those on 900 mg, 95% on 1200, 72% on 1500, and 78% on 1800 mg). In cohort 1, one pt died within a week without toxicity from P and 1 pt experienced grade 3 nausea, so this cohort was expanded. The other pts in the cohort had no toxicity. In cohort 4 one pt. took 300 mg daily for 6 days and was considered inevaluable. In cohort 3, grade 3/4 toxicity was not observed until after day 28 in 3 pts. Fifteen pts were evaluable for response; 1 had an unconfirmed partial response and 2 had stable disease. Conclusions: This study demonstrated that higher doses of P can be administered on a weekly schedule with divided doses. Weekly doses of 900 and 1200 mg lead to almost no toxicity in most pts. This study has been expanded to a phase 2 trial with direct comparison of weekly and daily doses. The grade 3/4 toxicities for each cohort are given in the table below.

[Table: see text]

[Table: see text]
A phase I study of axitinib (AG-013736), a potent inhibitor of VEGFRs, in combination with gemcitabine (GEM) in patients (pts) with advanced pancreatic cancer
Authors: Spano J. , Moore M. , Kim S. , Liau K. F. , Hee B. , Bycott P. , Boissaye P. , Francis S. , Peters D. , Rixe O.
Published: March 2020

13092

Background: Axitinib (AG-013736) is a novel small molecule inhibitor of the receptor tyrosine kinases with picomolar potency against VEGFR 1, 2 & 3 and nanomolar potency against PDGFR-beta and KIT. A phase I study in solid tumors identified 5 mg BID as the therapeutic dose; a phase II study in renal cell cancer demonstrated significant efficacy with a response rate (RR) of 46% (Rini et al, ASCO 2005). This study examined the safety, PK and preliminary efficacy of AG-013736 (AG) in combination with gemcitabine (GEM) as first-line therapy for advanced pancreatic cancer. Methods: A randomized phase II study was preceded by a phase I component. All patients (pts) in the phase I portion received 1000 mg/m2 GEM by 30-minute infusion on days 1, 8, and 15 followed by one week of rest from treatment. AG 5 mg p.o. BID was given beginning Cycle 1, Day 3 (C1D3). Eligible pts had no prior chemotherapy for advanced disease, ECOG 0–2, and no previous treatment with VEGF/VEGFR inhibitors, or anti-angiogenesis treatment. Full PK profiles were collected on C1D1 (GEM alone), C1D14 (steady state, AG alone), and C1D15 (GEM + AG). In the phase II trial, pts are randomized to AG or AG plus GEM beginning C1D1. Results: 8 pts were treated on the phase I portion of this trial. Toxicity: The primary Gr. 3/4 toxicity was hematologic: Gr. 4 anemia and Gr.3 thrombocytopenia in 1 pt and Gr. 3 neutropenia in 1 pt requiring a dose reduction for GEM in Cycle 3. Gr. 2 non-hematologic adverse events include pruritus (1 pt), abdominal pain (2 pts), epigastric pain (1 pt), melena (1 pt), and asthenia (2 pts). Gr. 2 hypertension was observed in 3 pts. Efficacy: Radiological assessment suggests 2 pts with partial response and 4 pts with stable disease: response assessments are ongoing. The median number of cycles is 3 [1,6]. Treatment for 4 pts is still ongoing: Cycle 6 (2 pts) and Cycle 2 (2 pts). Conclusions: This combination is safe and appears to be an effective treatment for advanced pancreatic cancer with significant tumor regression observed in 2 pts. Therapy was well tolerated with manageable toxicity. Additional investigation of AG-013736 in combination with GEM in the phase II setting for advanced pancreatic cancer is warranted.

[Table: see text]
Genetic polymorphisms of the adenosine triphosphate-binding cassette transporters (ABCC2, ABCG2) and irinotecan toxicity in cancer patients
Authors: Usui M. , Ando M. , Kitagawa C. , Ando Y. , Sekido Y. , Imaizumi K. , Shimokata K. , Hasegawa Y.
Published: March 2020

13096

Background: Irinotecan is subject to substantial interindividual variability in pharmacokinetic and the occurrence of unpredictably severe toxicities of leukopenia or diarrhea. These toxicities have been reported to be associated with increased levels of SN-38, the active metabolite of irinotecan. ABCC2 and ABCG2, members of adenosine triphosphate-binding cassette transporters, are involved in mediating the elimination of anionic antitumor drugs, such as SN-38 and SN-38G. Recently, Innocenti et al. reported SN-38G AUC and SN-38G/SN-38 AUC ratios were correlated with ABCC2 3972T>C. The variant ABCG2 421C>A was associated with low ABCG2 expression levels and altered sensitivity to several drugs, including SN-38, in vitro as compared with the reference-type protein. Methods: We assessed whether the variants ABCC2 3972T>C and ABCG2 421C>A would be associated with severe toxicity (leucopenia of grade 4 and/or diarrhea of grade 3 or worse) in 120 Japanese cancer patients in which 27 patients experienced severe toxicity. Results: 74 patients (62%) were homozygotes for the reference allele of ABCC2 3972T>C, 39 heterozygous (33%), and 7 homozygous (6%) for the variant, whereas 62 patients (52%) were homozygous for the reference allele of ABCG2 421C>A, 48 heterozygous (40%) and 10 homozygous (8%) for the variant. Logistic regression analysis did not show any significant associations between the occurrence of severe toxicity and carrying these variants (see Table ). Conclusions: It suggests that genotyping of ABCC2 3972T>C and ABCG2 421C>A would not be useful for predicting severe toxicity caused by irinotecan.

[Table: see text]

No significant financial relationships to disclose.
Synergistic inhibition of breast cancer cell lines with the combination of a dual inhibitor of EGFR/HER-2/neu and a Bcl-2 inhibitor
Authors: Witters L. , Witkoski A. , Planas-Silva M. , Viallet J. , Berger M. S. , Lipton A.
Published: March 2020

13100

Background: The epidermal growth factor receptor (EGFR; ErbB1) and HER-2/neu (ErbB2), members of the ErbB family of receptor tyrosine kinases, are overexpressed in a variety of human tumors and overexpression generally correlates with poor prognosis and decreased survival. Use of inhibitors of these receptors as monotherapies, e.g., trastuzumab, Iressa, and erlotinib, has led to advances in treatment, but many patients do not respond or develop resistance. The anti-apoptotic protein, Bcl-2, is also overexpressed in a number of human tumors. Inhibitors of Bcl-2 induce apoptosis and sensitize cancer cells to other therapies. This study assesses the effects of a combination of a reversible inhibitor of both EGFR and HER-2/neu that is similar to lapatinib (GW2974) and a pan inhibitor of the Bcl-2 family (GX15–070: Gemin X Biotechnologies, Inc.) on the growth of human breast cancer cells. Methods: The MCF-7 human breast cancer cell line transfected with a control vector, MCF/neo, and the HER-2/neu transfected MCF-7 cell line, MCF/18, were treated with various concentrations of GW2974 (0.25–10 μM) and/or the GX15–070 pan Bcl-2 inhibitor (50–500 nM). After a 3 day exposure, cell number was determined using the colorimetric MTT tetrazolium dye assay. Percent of control was normalized to corresponding concentrations of the solvent for both agents (DMSO). Results: Treatment with the GW2974 dual inhibitor or the GX15–070 pan Bcl-2 inhibitor resulted in dose-dependent growth inhibition in both the control and HER-2/neu transfected MCF-7 cell lines. The combination of both agents produced synergistic growth inhibition in both cell lines as confirmed by isobologram analysis. Conclusions: This study has demonstrated synergy with the combination of a dual inhibitor of EGFR and HER-2/neu and an inhibitor of Bcl-2 in control and HER-2/neu overexpressing MCF-7 human breast cancer cells. This finding warrants an evaluation of this combination in clinical trials for the treatment of patients with metastatic breast cancer.

[Table: see text]
Survivin silencing sensitizes imatinib-resistant CML cells to the cytotoxic effect of hydroxyurea
Authors: Stagno F. , Conte E. , Stella S. , Tirrò E. , Manzella L. , Palio E. , Giustolisi R. , Messina A. , Vigneri P.
Published: March 2020

13119

Background: Imatinib Mesylate (IM), a semi-specific inhibitor of the BCR-ABL tyrosine kinase, is currently the treatment of choice for Chronic Myeloid Leukemia (CML). However, about one third of CML patients treated with IM develop resistance to the drug because of reactivation of BCR-ABL kinase activity. This phenomenon is usually ascribed to the amplification of the BCR-ABL gene or to the selection of leukemic clones harboring point mutations that abrogate IM binding. To identify novel anti-apoptotic signaling pathways employed by BCR-ABL and devise strategies capable of killing IM-resistant CML cells, we investigated the interplay between BCR-ABL and the Inhibitor of Apoptosis Protein Survivin. Methods: Murine hematopoietic cells (32D) transduced with p210 BCR-ABL and human cell lines either positive (K562, KCL22, KYO1 and LAMA84) or negative (HL60) for the BCR-ABL oncoprotein, were analyzed for Survivin expression by western blot before and after IM treatment. Three different pathways (MAPK, PI3K and JAK2/STA3) potentially involved in BCR-ABL-mediated induction of Survivin were studied using inhibitors specific for each signaling cascade. The effect of Survivin on the proliferation and viability of IM-sensitive and IM-resistant CML cells was investigated after silencing Survivin expression with small interfering RNAs. Results: BCR-ABL tyrosine kinase activity induced an over-expression of Survivin in both human and murine hematopoietic cell lines. This over-expression was both at the transcriptional and the translational level and required the JAK2/STAT3 pathway. Survivin silencing by siRNA increased IM cytotoxicity in IM-sensitive cells but failed to restore IM efficacy in IM-resistant cells. However, Survivin silencing sensitized CML cells to the cytotoxic effect of hydroxyurea and enhanced the efficacy of this compound on three different murine cell lines are insensitive to IM because of point mutations in the BCR-ABL kinase domain (Ba/F3p210Y253F, Ba/F3p210D276G and Ba/F3p210T315I). Conclusions: Reduction of Survivin expression improves the efficacy of IM and increases the sensitivity of IM-resistant CML cells to hydroxyurea. Survivin may represent an attractive therapeutic target for both IM-sensitive and IM-resistant CML patients.

No significant financial relationships to disclose.