bluuebunny/crossref_metadata_2025 huggingface dataset:
Hypoxia effect on non-malignant prostatic urethra during prostatectomy
Authors: Josephson J. W. , Vira M. , Gillespie J. W. , Pinto P. A. , Srinivasan R. , Chuaqui R. F. , Emmert-Buck M. R. , Erickson H. S. , Coleman J. A.
Published: March 2020

20048

Background: With changes in technology and surgical technique paving the way to improved prostectomies, attention must be paid to the effects of hypoxia during variable surgical times. Hypoxia has been reported to play various roles that are implicated in the development of spurious molecular mutagenesis and tumorogenesis. However, the specifics leading to alterations and downstream effects in non-malignant tissues are unknown. Further, the consequence of hypoxia on tissues for analysis of tumor markers may be confounded. The objective of our study is to evaluate the effects of iatrogenic ischemia on surrounding non-malignant tissue in patients undergoing prostatectomy. As potential tissue markers to assess putative hypoxia tissue damage, gene expression levels in microdissected cell populations were evaluated. Methods: Normal prostatic urethra tissue was procured from five patients with laparoscopic 5 mm biopsy forceps at three time points during surgery and was immediately imbedded in OCT and placed on dry ice. First biopsy was taken prior to loss of blood supply after anterior bladder neck dissection. Second biopsy was taken directly after prostate was isolated from the patient. Third biopsy was taken ex vivo after vesico-urethral anastomosis. Urethral epithelium samples were normalized using the gold standard cell counting (2,000 microdissected cells) and quantitation of total RNA was conducted using NanoDrop technique. Gene expression analysis was assessed using quantitative RT-PCR (qRT-PCR) of four historically high and low abundant housekeeping genes (18s, ACTB, CYPA, and TFRC). Results: Housekeeping genes were up-regulated with an average four fold increase in transcript RNA at each time point respectively. Conclusion: These data suggest that hypoxia plays a putative significant role in the expression of genes in non-malignant tissues. The additional gene sets for analysis include traditional hypoxia related genes (e.g. HIF-1a, VEGF, AKT, and VHL) and non-hypoxia related genes (e.g. LAMP2, MTR, and STAT5B) to further investigate the prospective global up-regulation of gene expression. The conclusions may elucidate the role of hypoxia in disease processes and potentially its role in confounding or influencing tumor marker gene expression between patient samples.

[Table: see text]
Circulating levels of the breast cancer growth factor GP88 in the serum of breast cancer (BC) patients
Authors: Serrero G. , Tkaczuk K. , Tait N. , Golubeva O. , Dai H. , Feldman F. S. , Jones L.
Published: March 2020

20050

Background: The 88 kDa autocrine growth factor PC-Cell Derived Growth Factor (GP88) plays a critical role in breast tumorigenesis. GP88 expression was low in estrogen receptor positive cells, whereas in ER negative cells, it was constitutively overexpressed. Increased GP88 expression was associated with anti-estrogen therapy resistance in ER+ cells and Herceptin resistance in Her-2 overexpressing breast tumors. Antisense inhibition of GP88 expression in human breast adenocarcinoma lead to inhibition of tumor growth in vivo. Immunohistochemical studies have shown that GP88 was expressed in 80% of invasive ductal carcinomas in correlation with expression of poor prognosis markers whereas normal tissues and benign breast lesions were negative. Since GP88 is secreted by breast cancer cells, we examined whether GP88 was found in the circulation at an elevated level in the sera of breast cancer patients when compared to healthy individuals. Methods: A blood sampling study was conducted to determine the serum level of GP88 in healthy volunteers (HV) and breast cancer patients (BC pts). Ten ml of blood was drawn every three months to obtain serum. GP88 serum concentration was determined in triplicate by quantitative enzyme immunoassay using human GP88 as standard. 126 BC pts were accrued. . In addition, sera from 53 healthy volunteers were obtained to establish a GP88 baseline in HV. BC pts characteristics: race: Caucasian- 61, African American-60, Asian-5; median age: 52.5 (range 26–84), stage I-32, II-34, III-18, IV-42. Results: Circulating GP88 was measurable in the serum. Median level of GP88 was 32.8 ng/ml (range 15.3–42.8) in HV and 43.8 ng/ml (range 15.4–158.4) in BC pts, (p-value = 0.0007). Conclusions: GP88 is measurable in the sera of HV and BC pts. Comparison between the two groups indicates that GP88 level is significantly higher in the sera of BC pts. These studies are important since it identifies GP88 as a measurable biomarker that is also a therapeutic target of malignant transformation or malignant progression of breast carcinoma (BC). Future studies will examine the correlation of GP88 level with BC prognostic factors. Correlation between the serum level of GP88 and therapeutic response to systemic therapy in breast cancer patients will also be assessed.

[Table: see text]
geNorm assessment of the efficacy of novel reference genes ALG9, TFCP2 and ZNF410 compared to GAPDH for real time normalization in GI carcinoids
Authors: Eick G. , Kidd M. , Mane S. , Nadler B. , Champaneria M. , Pfragner R. , Modlin I. M.
Published: March 2020

20052

Background: Robust quantitation of potential clinical marker genes using quantitative real-time PCR (Q RT-PCR) is critically dependent on accurate normalization. Although GAPDH has historically been used for normalization, its expression has been shown to vary widely between different tissues and experimental conditions. Additionally, conventional normalization strategies based on a single housekeeping gene can lead to large normalization errors. The determination of a panel of genes that have robust expression in the experimental system being studied is therefore essential to ensure accurate normalization and interpretation of results. Methods: Based upon the availability of large-scale gene databases, we developed methodology to identify highly expressed genes (mean log-transformed expression levels: 4–8 in all samples) with low variability (S.D. < 0.22) in a Affymetrix U133A dataset of 36 gastrointestinal tumors and normal tissues. Eight novel candidate reference genes were identified and their expression levels established by Q RT-PCR in an independent set of GI tissue samples (n = 24). The geNorm tool was used to identify the most stably expressed set of genes amongst the 8 candidate genes. The expression levels of 3 potential GI tumor marker genes, namely the adhesin MAGE-D2, the metastasis-associated MTA1, and the mitosis regulator, NAP1L1, were normalized to GAPDH or geNorm and compared. Results: geNorm identified 3 genes, ALG9, TFCP2 and ZNF410, as the most robustly expressed control genes. GAPDH, in contrast, exhibited the highest variability and was considered the least stable gene of the nine evaluated. Two previously-identified target genes, MAGE-D2 and MTA1, were significantly elevated (p < 0.05) in malignant tumor samples (vs normal GI samples) when normalized by geNormATZ but not when normalized using GAPDH. NAP1L1 was only over-expressed in small intestinal carcinoids (normalized to geNormATZ). Expression levels of this gene were high in normal gastric mucosa. Conclusions: We provide a robust basis for the establishment of a reference gene set using GeneChip data and provide evidence for the clinical utility of identifying reference genes using the geNorm approach in GI neuroendocrine tumors.

No significant financial relationships to disclose.
What is the role of QTc prolongation assessment in new drugs development phase I oncology trials?
Authors: Curigliano G. , Cipolla C. , Sessa C. , Noberasco C. , De Pas T. , Colombo N. , Spitaleri G. , Putzu C. , Milani A. , De Braud F.
Published: March 2020

2006

Background: QT interval prolongation associated with “torsades de pointes” (TdP) has been a common cause of withdrawal from the market for several promising drugs. We determined the degree of QT prolongation in patients treated within a phase I study with a thioxanthone derivative known to have arrhythmogenic potential. Methods: Clinical data and serial ECGs from 31 patients with advanced tumors who received 86 courses of investigational drug were prospectively reviewed. The drug has been administered intravenously over 24 hours every 3 weeks. Patients have been on a 24 hour Holter monitor until 24 hours after infusion was complete. Three baseline ECGs were done and compared to those every 6 hours during therapy and once 6 hours after the infusion was complete. ECGs were read at a central lab according to a standard protocol. All QT measurements were then corrected for heart rate (QTc) using Bazett’s formula (QTc = QT interval divided by the square root of the R-R interval). Results: Overall,843 ECG tracings were obtained, all evaluable for analysis. No basal ECG showed significant abnormalities. Prolonged QT intervals developed in 2 patients without clinical symptoms (1 patient had intervals 500 milliseconds).In both cases it was associated with the maximum concentration of the drug. Compared with baseline, the QTc interval was prolonged by 30 to 60 milliseconds in 20% of total tracings, and by more than 60 milliseconds in 2% of ECGs. In patients receiving multiple courses, QTc intervals returned to pretreatment levels before the second course. Conclusions: The assessment of QTc prolongation was a major effort in this study but produced an accurate information about such event. In phase I study such an effort is justified when arrhythmogenicity is suspected. The timing of collection of ECGs should be guided by the available preclinical information about the pharmacokinetic profile of the drug. Nevertheless uncertainty remains regarding the specific relationship between the degree of QT prolongation and the risk of life-threatening arrhythmias. The decision to use the drug ultimately has to be based on an estimation of the perceived risk relative to expected benefits for patients.

No significant financial relationships to disclose.
Survival in women with breast cancer having children with pediatric tumors—-A nationwide cancer register study
Authors: Olsson H. , Bladström A.
Published: March 2020

20071

Background: Breast cancer is a heterogenous disease with a minoruty of cases having a hereditary background. Hereditary factors could affect disease prognosis and it has been postulated that breast cancer patients having p53 germline mutations have an especially bad prognosis. In the present study survival has been compared in women with breast cancer with and without having a child with a pediatric tumour. Methods: Using and linking the population based fertility registry and cancer registry the offspring of all women with breast cancer diagnosed between 1961–1999 was traced in Sweden. All women with breast cancer with a pregnancy were included. Further all childhood cancer cases < 21 years of age were identified among the offspring. Likewise using census data the women’s overall survival was followed. Results: Patients with breast cancer having a child with a childhood malignancy (n = 254) had a significant worse survival compared with other women with breast cancer (n = 74,781), RR = 1.25, 95% CI 1.02–1.55, p < 0.04, adjusted for age at diagnosis, year of diagnosis, parity and time since last pregnancy. Mothers with children with pediatric sarcomas or acute myeloid leukemias seemed to be strongest associated with a worse survival in the mother (RR = 1.38 and RR = 1.69). The worse survival of the mother was present regardless if the breast cancer diagnosis was before or after 50 years of age. Conclusion: The results suggest that genetic syndromes such as the Li-Fraumeni syndrome and possibly other childhood/adult cancer syndromes if present worsens the survival for breast cancer patients. This familial factor may be of importance for future treatment stratification. It is unknown whether other tumor diseases in the mother may be associated with a worse survival if their children have had cancer.

No significant financial relationships to disclose.
Validation of clinical prognostic factors for survival in 1,043 Japanese patients with metastatic renal cell carcinoma (mRCC)
Authors: Tomita Y. , Naito S. , Ozono S. , Terachi T. , Naito S.
Published: March 2020

20075

Background: Increasing number of patients and death of RCC has been reported in many countries including US and Japan. For choosing optimal treatment, determining risk of patients is essential. Although several prognostic factors of mRCC has been investigated, it was based on data of Caucasian. Since racial difference in biological character of cancer has been identified in several types of cancer, we validated the relationship between reported prognostic factors and survival in patients with mRCC in Japanese patients. Methods: We reviewed the medical records of 1,043 patients who had mRCC in the period to follow up, or at the first visit examination between 1988 and 2001 of 26 center in Japan. Results: Average survival of the 708 patients who died for renal cell carcinoma was 20.84 months. On the other hand, histological high grade (> WHO grade 2), metastasis at the first visit examination, short period from the first visit to recognition of metastasis (within 12 months), brain, bone or, liver metastasis, a number of metastatic sites (>1), high performance status (ECOG PS >0), high corrected serum calcium, low hemoglobin, and high CRP were prognostic factors for survival. However, LDH were not related. The median time to death with zero risk factor was 38.5 months, but those with >3 risk factors was 10 months. The patients with >4 risk factors was 4 months. Conclusions: Almost all prognostic factors reported in previous studies mainly based on Caucasian data had same impact on Japanese patients with mRCC. However, elevation of LDH did not have relationship to prognosis. Moreover, the time to death from diagnosis of mRCC in patients with poor prognostic factors was likely shorter compared with other studies. These results suggest Japanese with mRCC might have different character from those in US or European countries though further study in patient’s and tumor characteristics is warranted.

No significant financial relationships to disclose.
Discordence of HER-2/neu overexpression and estrogen receptor between the primary breast cancer and axillary lymph node with metastasis
Authors: Shahid R. K. , Ahmad I. , Yadav S. , Sami A. , Haider K. , Rees H. , Ahmed S.
Published: March 2020

20106

Background: Adjuvant hormonal therapy and trastuzumab are known to be associated with a significant reduction in cancer-related mortality in a subset of women with breast cancer. The treatment decision is usually based on immunohistochemistry (IHC) staining of the primary tumor. We reported here a case of discordance of HER-2/neu overexpression and estrogen receptor status between the primary breast cancer and axillary lymph node with metastasis. Methods: IHC and fluorescence in situ hybridization (FISH) analysis was perfromed for the assessment of HER-2/neu. E-cadherin staining was done on the primary tumor and lymph node with metastasis. Results: A 52 year old woman presented with right breast lump. The imaging studies showed a 5 × 5 cm speculated mass. No other abnormalities were noted. An excision biopsy revealed 3 cm moderately differentiated invasive lobular carcinoma. Subsequently a wider excision and axillary lymph node dissection was done. 2/20 lymph nodes were involved by the cancer. IHC was done in the primary tumor and the lymph node with metastasis. IHC of the primary tumor revealed less than 1% nuclei positive for estrogen receptor (ER), 0% nuclei positive for progesterone receptor (PR), and no HER-2/neu overexpression. IHC of the lymph nodes with metastasis revealed 80% nuclei positive for ER, less than 1% nuclei positive for PR, and equivocal HER-2/neu overexpression. FISH analysis for HER-2/neu gene was done on the primary tumor which revealed no amplification in the tumor nuclei (Her2/CEP17 ratio was 3.6/2.7 = 1.3) whereas FISH analysis on the lymph node section revealed amplification in tumor nuclei (Her2/CEP 17 ratio of 7.3/1.9 = 3.9). E-cadherin staining of the primary breast tumor as well as the lymph node with metastasis revealed no membrane staining of the malignant cells in either section. Conclusions: The unusual occurrence of a divergent IHC profile and FISH analysis results between the primary carcinoma and metastatic carcinoma may be due to a subselection of a malignant clone which were not represented in the section of primary tumor chosen for IHC. Future studies are required to determine the utility of IHC staining of metastatic cells involving the lymph nodes if primary tumor is negative for HER-2/neu & ER.

No significant financial relationships to disclose.
Mystery of tumor specific immunity: LIGHT turns on silent tumor specific immunity
Authors: Cao L. T.
Published: March 2020

20107

Background: More than 2000 tumor antigens have been identified so far. However, none of these tumor antigens are approved to be immunogenic. Genetic alteration which activates uncontrolled cell growth transforms normal cells to tumor cells. Tumor cells, therefore, do not carry any foreign antigens and thus tumor antigens are not immunogenic (unless in the presence of viral infection). The so-called tumor specific immunity is actually the host versus graft effect, not specific for tumors. Methods and Results: Using proper controlled tissue graft as control, our previous data indicated that the hosts could accept both the graft and the tumor. The immune system is silent to tumor antigens. By transfecting tumor cells with a modified LIGHT (Lymphotoxins, shows Inducible expression, and completes with herpes simplex virus Glycoprotein D for Herpes virus entry mediator, a receptor expressed by T lymphocytes) expressing vectors, Dr. Fu at the University of Chicago is able to eradicate tumors in his animal model. The expressed LIGHT on the tumor cell surface serves as tether for the naïve T cells and the stromal cells by binding to lymphotoxin-beta receptor on stromal cells and herpes virus entry mediator on T cells. The trio of tumor-LIGHT-stroma working like a foreign antigen attracts naïve T cells and increases the infiltration of naïve T cells within the tumors. The trio then co-activates naïve T cells to destroy tumors. The LIGHT mechanism may be similar to that in autoimmune phenomenon. Conclusion: Tumor specific immunity may be silent; however we can modify tumor antigens and activate silent immune system to destroy tumors. We currently have four immunotherapies against cancer. The LIGHT approach or in situ co-stimulation therapy, vaccine therapy, and ex vivo cell-transfer therapy are three active immunotherapies. The fourth one is a passive immunotherapy: the near-specific, personalized immunotherapy.

No significant financial relationships to disclose.
Fraction of LOH on chromosome 17 and 18 (LOH-ratio) predicts colorectal cancer outcome
Authors: Kanazawa T. , Watanabe T. , Nagawa H.
Published: March 2020

20114

Background: Identification of patients at high risk for recurrence remains a central issue in the treatment of colorectal cancer. Our goal was to identify predictive factors for recurrence in colorectal cancer patients. Methods: DNA from 84 colorectal cancers were analyzed for wide-ranging allelotyping. Using 27 microsatellite markers spanning every 10cM on chromosome 17 and 18, we defined the LOH-ratio as the proportion of markers which show LOH out of 27 markers. Tumors were grouped into two groups by the median value of LOH-ratio (0.19). Recurrence free survival was compared with Kaplan-Meier analysis and log rank statistic. The Cox proportional hazards regression model was used for both univariate and multivariate analysis of recurrence free survival. Results: Log rank statistic revealed that LOH-ratio, stage, lymph node metastasis significantly related to recurrence free survival. On univariate analysis, significant predictors of Recurrence free survival were LOH-ratio, lymph node metastasis, Dukes’ classification, and pathological type. On multivariate analysis, LOH-ratio (HR 3.1, p = 0.02) and lymph node metastasis (HR 5.2, p = 0.002) independently predicted for recurrence free survival. Conclusions: LOH-ratio and lymph node metastasis were the only independent predictors of recurrence free survival. Altogether with lymph node metastasis, LOH-ratio could help to improve postoperative surveillance and adjuvant therapy.

No significant financial relationships to disclose.
A phase I trial of weekly combination KOS-862 (Epothilone D) and trastuzumab in HER-2 overexpressing malignancies
Authors: Cortes J. , Climent M. A. , Gomez P. , Ruiz A. , Calvo E. , Zhang X. , Perez E. S. , Demario M. , Baselga J.
Published: March 2020

2028

Background: KOS-862 (Epothilone D) (K) has shown single agent activity against metastatic breast cancer (MBC) and NSCLC. Trastuzumab (H) is standard therapy for HER-2 overexpressing MBC. This phase IB trial determined the safety and PK profile of KOS-862 in a weekly schedule with H. Methods: Eligibility: pts with HER-2 overexpressing (IHC 3+ or FISH +) chemo-refractory tumors, ECOG ≤ 2, neuropathy ≤ gr 1, no CNS metastases, and no H ≤ 6 mo. prestudy. H given as a 4 mg/kg loading dose, then 2 mg/kg wkly. K given as a 90 min iv infusion wkly × 3 Q 4 wks after H. K dose escalation cohorts: 70, 85 & 100 mg/m2. H1 + H2 antagonists + corticosteroid prophylaxis given 30–60 min prior to K. DLT: gr 4 hematologic or gr 3 other toxicity, or toxicity requiring >4 wk treatment delay. Cycle 1 + 2 PK samples obtained for K + H. Results: 13 pts treated from 9/04 - 8/05. Median age 51 yrs (36–65). All pts had MBC. Prior H therapy n = 9, prior taxane n = 11. Enrollment: 70 mg/m2 (4), 85 mg/m2 (3), and 100 mg/m2 (6). Mean number cycles 3.9 (1–6). The 100 mg/m2 cohort was expanded to 6 pts per study design; no DLTs were noted. Toxicity: Principal neurotoxicities (gr 3 noted, if present): parasthesias (10 pts, 1 gr3), neuropathic pain (5 pts, 2 gr3), dizziness (4 pts), dysguesia (4 pts), ataxia (3 pts), transient confusion (3 pts), dysesthesias (2 pts), insomnia (2 pts). No gr 4 neurotoxicities reported. Non-neurologic toxicities were limited to gr 1/2 and included diarrhea, pyrexia, abdominal and back pain, arthralgias. PK (n = 13): KOS-862 CL 21 ± 10 L/hr, Vz 260 ± 110 L, T1/2 9.8 ± 3.9 hr. Seco-metabolite was 67% ± 35% of K. At 100 mg/m2 K dose (n = 6): Cmax 3.3 ± 1.1 μg/mL and AUCinf 11.0 ± 7.5 h*(g/mL). Neither K nor its seco-metabolite PK were affected by H. Responses: An 85 mg/m2 K pt (no prior H or taxane) had confirmed PR and a 100 mg/m2 K pt (prior taxane) had an unconfirmed PR. A 2nd 100 mg/m2 pt (no prior H or taxane, bone only disease) had > 90% CEA reduction. Conclusions: This phase I study demonstrates feasibility of full dose wkly KOS-862 coadministration with wkly H. Treatment-related toxicities were all gr 1 / 2, except for 3 cases gr 3 paresthesias and neuropathic pain. PK of K and its principal metabolite were not affected by H. Three clinical responses were observed; a phase II trial of this combination is ongoing in minimally-pretreated MBC.

[Table: see text]
Phase 1 and pharmacokinetic study of weekly docosahexaenoic acid-paclitaxel (Taxoprexin) in resistant solid tumor malignancies
Authors: Fracasso P. M. , Govindan R. , Ellis M. J. , Tan B. R. , Linette G. P. , Picus J. , Goodner S. A. , Jones S. , Bellet R. E. , Burris H. A.
Published: March 2020

2036

Background: Docosahexaenoic acid-paclitaxel (DHAP) is a covalent conjugate of paclitaxel and the essential fatty acid, docosahexaenoic acid. Preclinical studies have demonstrated increased activity relative to paclitaxel with the potential for an improved therapeutic ratio. We performed a phase 1 and pharmacokinetics (PK) study of DHAP in patients with resistant solid tumor malignancies. Methods: Patients were treated with DHAP administered by 2-hour intravenous infusion weekly for three out of four weeks. Dose escalation started at DHAP 200 mg/m2 and was increased by 100 mg/m2 per dose level ending at 600 mg/m2. Blood samples for PK of DHAP and paclitaxel derived from DHAP were collected in cycle 1 at 0 (prior to the start of the infusion), 2 (prior to the end of the infusion), 3, 4, 9, 26 hours and just prior to the next weekly treatment on days 8, 15 and 29. Results: Twenty-one patients enrolled and received 39 cycles of treatment (mean = 2). Three patients received DHAP at 200 mg/m2 (dose level 1), four at 300 mg/m2 (dose level 2), three at 400 mg/m2 (dose level 3) and 500 mg/m2 (dose level 4) each, and eight at 600 mg/m2 (dose level 5). In cycle 1, grade 2/3 neutropenia resulted in treatment delays for 8 patients, 6 of whom received treatment at dose level 4 and 5. Two of the 8 patients, treated with DHAP 600 mg/m2, went on to experience grade 4 neutropenia lasting < five days on later cycles. Only one patient had grade 2 sensory neuropathy, which occurred after 3 cycles of treatment. Preliminary PK analyses on all patients demonstrated that there is not a substantial accumulation of DHAP or paclitaxel with weekly treatment. Detailed PK analyses will be presented. Eighteen of the 21 patients were evaluable for response. Three patients, one each with esophageal and lung carcinoma and melanoma, had stable disease for 11, 16, and 17 weeks, respectively. Conclusions: DHAP administered weekly at 600 mg/m2 was well-tolerated and the maximum dose administered in this study. The slow release of paclitaxel from DHAP and the weekly schedule approximates continuous infusion paclitaxel which should be more active than every 3 week or weekly taxanes. Further studies with DHAP in non-small cell lung carcinoma and melanoma are on-going.

[Table: see text]
Phase I dose-escalating study of bi-weekly pemetrexed (MTA) with fixed dose rate gemcitabine (gem) in patients with advanced cancer
Authors: Chen A. L. , Ryan T. , Muggia F. , Chachoua A. , Escalon J. , Moore S. , Haniff F. , Levinson B. , Nicol S. , Hochster H.
Published: March 2020

2049

Background: Fixed dose rate (FDR) gem may optimize the documented benefit of gem in pancreatic, NSCL, head and neck, and breast cancers. Preclinical data of the combination of MTA and gem indicate synergy. We studied a bi-weekly combination of MTA and FDR gem to improve tolerance over the previous “Day 1, 8” regimen. Methods: MTA followed by FDR gem infusion was given every 14 days in a dose-escalation design below. Dose-limiting toxicities (DLT) is defined as: grade 4 neutropenia ≥ 5 days, febrile neutropenia, grade 4 thrombocytopenia, or grade 3–4 non-hematological toxicities in 1st cycle. 27 patients were enrolled with median age 59 (range 41–82), males/females 18/9, and ECOG PS 0/1 (5/22). All received Vitamin B12 (1 mg IM prior to study & q9 weeks) and folic acid (350–1000 mcg po qd) supplementation 1–2 weeks prior and during study plus dexamethasone 4 mg po BID day before, of, and after chemotherapy. Results: Dose level 7 has been reached and is being expanded with DLT in 1 patient. Toxicities per patient during all cycles include grade 3–4 neutropenia (8/27 patients), grade 3 anemia (3), brief febrile neutropenia (6), grade 3 rash (1 patient), grade 3 low back pain (1), grade 3 renal failure (1), grade 5 perforated duodenal stent (1). In 19 patients evaluable for response, there were 11 stable disease and 8 progression of disease. Conclusion: Recommended phase II dose for bi-weekly MTA with FDR gem is likely to be 800 mg/m2 and 1,200 mg/m2 × 120 minutes. This regimen allows full doses and dose intensity of both drugs to be administered on a simple schedule with excellent tolerance. A phase II study in biliary cancer is planned. This study is supported in part by Eli Lilly.a This toxicity was at baseline, therefore not drug related.b, c These toxicities were in one patient and were not drug related, rather related to the individual patient’s status going into therapy.

[Table: see text]

[Table: see text]
Pharmacogenomic analysis for individual response to CPT-11 in colorectal cancer: Prediction formula of tumor response using novel marker genes and genotypes associated with the toxicity
Authors: Narahara H. , Sugimoto N. , Tomita N. , Murata K. , Fukunaga M. , Ohue M. , Sakaeda T. , Ishida H. , Tanimoto K. , Nishiyama M.
Published: March 2020

2060

Background: Despite recent encouraging data, the clinical efficacy and toxicity of CPT-11 in most colorectal cancer patients remain unpredictable. We conducted this prospective study to develop a prediction formula of efficacy using expression data of genes newly identified in vitro and to evaluate the clinical significance of several genetic polymorphisms known as potent predictors of toxicity. Methods: CPT-11 was intravenously administered on Days 1, 8, and 15, every 4 weeks in chemo-naive patients with stage IV colorectal cancer after palliative operation. Tumor samples were collected at surgery and PK analysis was done on day 1 of cycle 1. Tumor response and toxicity were evaluated by RECIST and CTCAE, respectively. Results: Forty seven pts were enrolled. All pts were assessed for tumor response, toxicity, gene expression, genotype, and PK. Partial response was obtained in 11 cases, and common grade 3/4 toxicities were neutropenia (15%), leucopenia (5%), and diarrhea (4%). Despite no correlation of UGT1A1*28 (hetero=5, homo=1) alone with the toxicities, there were associations between UGT1A1*28 or *7 (hetero=1) with UGT1A1 -64G>C (hetero=3) and G3/4 leucopenia (p=0.036). UGT1A1 1091C>T (hetero=3) was found to be associated with leucopenia (p=0.0009), neutropenia (p=0.012), and diarrhea (p=0.036). UGT1A1 1091C>T, ABCC2 -24C>T and 3972C>T were correlated with SN-38 AUC. We identified 7 novel potent marker genes including AMD1, CTSC, and EIF1AX for CPT-11 efficacy in vitro, through 2 different microarray analyses and subsequent real-time RT-PCR. We then successfully developed the best linear model, which converted the quantified expression data into objective tumor response, using 18 data sets of gene expression and clinical response. Utility-confirmation analyses using other clinical samples appeared to show that the formula could accurately predict tumor response (r=0.712, p=0.042). Conclusions: Polygenetic strategies using several known polymorphisms for toxicity and a prediction formula using 7 novel genes for efficacy would be of predictive value for individual response to CPT-11.

No significant financial relationships to disclose.
Plasma levels of SN-38 after co-administration of gefitinib and oral irinotecan in young and old mice
Authors: Cook D. R. , Merz A. L. , Zirrolli J. A. , Basche M. , Gustafson D. L.
Published: March 2020

2062

Background: Preclinical studies demonstrate that irinotecan, approved for I.V. administration, is most effective when administered at frequent, low doses. This administration schedule would be most practical if irinotecan were administered orally. However, the oral availability of irinotecan is limited. Gefitinib can alter the oral bioavailability of irinotecan through inhibition of gut transporters (ABCG2) that prevent intestinal absorption. Thus, we evaluated a potential method for giving irinotecan at low doses on a protracted schedule by administering orally with gefitinib. Additionally, we hypothesized that the pharmacokinetics of irinotecan may differ in older versus younger adult mice. Plasma levels of SN-38 were measured and compared in old and young Balb/c mice given oral irinotecan alone or with gefitinib. Methods: Adult (2 months old) and older, retired breeder (10 months old) Balb/c mice were orally administered irinotecan at 10 mg/kg, 20 mg/kg or 40 mg/kg either alone (control) or with 30 mg/kg PO gefitinib daily for 5 days. On the fifth day, the mice were sacrificed approximately 4–6 hours after administering drugs. Plasma was collected and SN-38 levels determined by HPLC. Results: Nearly all mice receiving irinotecan with gefitinib had a 3 fold or more increase in the level of SN-38 in the plasma as compared to the mice receiving irinotecan alone. The older mice had a more dramatic increase. Conclusions: The plasma levels of SN-38 were consistently elevated in mice treated with oral gefitinib and irinotecan when compared to mice treated with irinotecan only. The more pronounced increase in SN-38 levels seen in the older mice may be due to changes in drug elimination with aging. The results indicate that oral irinotecan can be given in the presence of gefitinib resulting in higher levels of SN-38 in the plasma of young and older mice. Combination treatment allows easier titration of irinotecan in protracted dosing protocols evaluating efficacy and toxicity in older mice, and ultimately, humans.

[Table: see text]

No significant financial relationships to disclose.
Autoresistance to ErbB2 kinase inhibitors: Elucidating mechanisms and identfying strategies to prevent its onset in breast cancer
Authors: Xia W. , Bacus S. , Hedge P. , Husain I. , Strum J. , Liu L. , Paulazzo G. , Harris J. , Trusk P. , Spector N.
Published: March 2020

2075

Background: Clinical efficacy of ErbB1/ErbB2 kinase inhibitors is limited by the development of acquired autoresistance. Developing clinically relevant models to study the mechanism(s) of acquired autoresistance will identify therapeutic strategies to prevent its onset. Methods: BT474 cells, an ErbB2-overexpressing/ER+ breast cancer line sensitive to lapatinib-induced apoptosis were chronically exposed to lapatinib, simulating chronic administration of lapatinib in the clinic. Lapatinib-resistant cells (rBT474) and single cell clones were established. Baseline gene expression in parental BT474 and rBT474 cells was compared using Human Affymetrix oligonucleotide arrays. Sequential tumor biopsies were analyzed by quantitative IHC as part of a clinical trial using lapatinib monotherapy (1500mg/d) in patients with metastatic breast cancer. Results: Gene expression and protein analysis indicates that acquired resistance to lapatinib is mediated by increased estrogen receptor (ER) signaling rather than loss of ErbB2 expression or insensitivity of the ErbB2-MAPK-PI3K pathways to lapatinib. Activation of ER signaling results from de-repression of FOXO3a and transient up-regulation of caveolin-1, both facilitating ER transcriptional activity. Gene interference studies confirm that acquired resistance involves a switch in survival dependence and regulation of survivin from ErbB2 to ER. Since ErbB2 signaling is operative in rBT474 cells, treating parental BT474 cells with lapatinib plus (i) fulvestrant, or (ii) estrogen deprivation prevents the onset of acquired lapatinib resistance compared with either therapy alone. Importantly, increased ER signaling occurs in patients with ErbB2-overexpressing/ER+ breast cancers treated with lapatinib monotherapy, consistent with changes in cell lines. Conclusions: Chronic lapatinib monotherapy induces a shift in the dependence of cell survival from ErbB2 to ER. Combining lapatinib with selected anti-estrogens prevents its onset in cell lines and warrants clinical validation. Establishing models of acquired resistance simulating the clinic provides a tool to identify strategies to improve treatment options for patients with ErbB2-overexpressing breast cancers.

[Table: see text]
Phase I dose-escalation study of ZK 304709, an oral multi-target tumor growth inhibitor (MTGI), administered for 14 days of a 28-day cycle
Authors: Ahmed S. , Molife R. , Shaw H. , Steward W. , Thomas A. , Barrett M. , Kowal K. , McCoy C. , De-Bono J.
Published: March 2020

2076

Background: ZK 304709 is a novel oral MTGI that induces cell cycle arrest and inhibits tumour angiogenesis by selectively inhibiting Cyclin Dependent Kinases (CDKs) 1, 2, 4, 7 and 9,VEGF-R 1, 2 and 3, and PDGF-Rβ tyrosine kinases. Methods: Adult patients (pts) (WHO PS ≤2) with a histologically or cytologically confirmed solid tumor, resistant or refractory to conventional therapy, were eligible. ZK304709 was administered orally, once daily, at a 15 mg starting dose, on days 1–14 of a 28-day cycle, then escalated by 33–100% depending on incidence of drug-related toxicity ≥ grade (gr) 2 (CTC v2.0). At least 3 pts were treated at each dose level. The primary objective was to identify the maximum tolerated dose (MTD) and dose-limiting toxicities (DLT). Secondary objectives were to determine the tolerability, pharmacokinetic (PK) profile, and preliminary efficacy. Results: Interim results were available for 24 pts (19 M/5 F; median age 56.5) at 5 dose levels (15–120 mg qd). Pts received a median of 2 cycles (range 0–10). Common drug related toxicities were nausea, vomiting, and fatigue. Two DLT were observed: dizziness and hypertension. However, the MTD has yet to be established. The PK profile for dose levels up to 90 mg demonstrated rapid absorption and a dose-dependent increase of exposure and Cmax. Disease stabilization for ≥4 cycles has been observed. Conclusions: ZK 304709 was rapidly absorbed and has been tolerated on this schedule at up to 120 mg qd. The MTD has not been reached and enrolment is ongoing. These preliminary data demonstrate that oral delivery on this schedule of an agent that inhibits both cell cycle and angiogenesis is feasible.

[Table: see text]
Combination immunotherapy with rituximab and local delivery of CpG oligonucleotide in a syngeneic murine model expressing human CD20
Authors: Hurvitz S. A. , Betting D. , Kafi-Tehrani K. , Golay J. , Olafsen T. , Wu A. M. , Timmerman J. M.
Published: March 2020

2502

Background: Immunostimulatory agents such as CpG oligonucleotides (CpG) may improve the efficacy of rituximab (R) by augmenting antibody-dependent cellular cytotoxicity. We have generated a murine B cell lymphoma expressing human CD20 (38C13-huCD20) for growth in syngeneic wild-type (WT) mice to efficiently study combination immunotherapy in an immunocompetent model. Methods: Groups of mice were inoculated s.c. with tumor cells and after tumor establishment, were treated with R on days 1 and 3, and either intratumoral (i.t.) or systemic (intraperitoneal, i.p.) CpG days 1, 2, 3, 5, 7 and 9. Tumor size was monitored daily and animals sacrificed when tumors reached 1.4 cm. Results: 38C13-huCD20 tumors were shown to express high levels of CD20 by flow cytometry, immunohistochemistry and microPET. While single agent R prevented growth of 100% of tumors when given on the day of inoculation, therapy of established tumors with either R or CpG alone provided no cures. In contrast, R plus i.t. CpG resulted in complete regressions (CR) of tumors in some mice. The route of CpG delivery was critical as only i.t., not systemic (i.p.) CpG combined with R led to CRs. All mice with CR survived challenge with a lethal dose of WT tumor cells, suggesting induction of immunity against non-human CD20 tumor antigens. Cyclophosphamide (Cy) was used to cytoreduce bulky tumors prior to treatment with R/CpG. Cy alone resulted in initial tumor shrinkage but no cures. In this stringent setting, there were no survivors after Cy/R. Long term tumor-free survivors were seen only in mice receiving i.t. CpG ± R. Addition of i.t. CpG significantly improved survival over Cy/R (p=0.018), while addition of i.p. CpG did not (p=0.24). Adding a second cycle of R/CpG i.t. on day 14 after Cy further improved survival (day 55) compared to 2 cycles of R alone (p=0.01) or 2 cycles of i.t. CpG alone (p=0.002). Conclusions: We have identified a novel combination immunotherapy resulting in eradication of an aggressive B cell lymphoma. Admininstration of CpG into the tumor bed was critical in enhancing the R effect. Ongoing studies are evaluating underlying immunologic mechanisms. Such models should allow identification of promising strategies appropriate for translation into clinical trials.

[Table: see text]
Interleukin-21 (IL-21): Tolerability and anti-tumor activity following two 5-day cycles in patients with stage IV melanoma (MM) or renal cell carcinoma (RCC)
Authors: Thompson J. A. , Curti B. D. , Redman B. G. , Weber J. S. , Agarwala S. S. , Sievers E. L.
Published: March 2020

2505

Background: IL-21 enhances NK cell activity and the proliferation and activation of CD8+ T cells. A Phase 1 outpatient dose-escalation trial of IL-21 in patients with MM or RCC established a maximum tolerated dose and demonstrated biologic effects as measured by increased soluble CD25 levels and decreased lymphocyte counts. Partial responses (PR) were observed in 2 patients with RCC (J Immunother. 2005; 28:641–642). Methods: IL-21 was administered by intravenous push at 30 μg/kg/day for two 5-day cycles separated by a 9- or 16-day rest to a total of 34 patients (18 MM and 16 RCC) to further characterize safety and evaluate preliminary signs of anti-tumor activity. Results: All patients received 2 cycles of treatment with the exception of one patient who discontinued secondary to pleural effusion. Common drug-related Grade 1–2 adverse events (AEs) included pyrexia (n=27), fatigue (n=25), chills (n=18), headache (n=18), and rash (n=16). Related Grade 3 AEs included pleural effusion, abdominal pain, hypophosphatemia, and thrombocytopenia (n=1 each). Laboratory abnormalities commonly observed were decreased lymphocytes (n=31), platelets (n=31), albumin (n=29), and hemoglobin (n=27); increased triglycerides (n=27) and ALT (n=26). Except for lymphopenia (an expected pharmacodynamic response), most toxicities were Grade 1–2. AEs and laboratory abnormalities were generally transient. Final study results, including response data, will be presented. To date, of 28 patients evaluable for response (i.e., those who received at least 8 of 10 doses and have had a final evaluation), one PR has been confirmed in a patient with MM and 18 patients have stable disease two months following initiation of therapy. Conclusions: Two 5-day outpatient cycles of IL-21 at 30 μg/kg/day have been reasonably well tolerated and have demonstrated preliminary evidence of anti-tumor activity in patients with advanced MM or RCC.

[Table: see text]
A phase II trial of denileukin diftitox to treat refractory advanced-stage ovarian cancer
Authors: Barnett B. G. , Ruter J. , Brumlik M. J. , Kryczek I. , Cheng P. J. , Zou W. , Curiel T. J.
Published: March 2020

2506

Background: Denileukin diftitox is a fusion toxin consisting of interleukin-2 genetically fused to diphtheria toxin. It is approved to treat cutaneous T cell lymphoma. Our recent Phase I trial demonstrated that denileukin diftitox depletes human regulatory T cells (Tregs), and is associated with improved measures of T cell immunity and clinical improvements in ovarian cancer. This Phase II trial tests whether denileukin diftitox has clinical activity in the treatment of relapsed epithelial ovarian cancer. Methods: Patients with Stage III or IV ovarian cancer who have failed first-line therapy with optimal surgical debulking and platinum-based chemotherapy were eligible. Denileukin diftitox was given at 12 mcg/kg once monthly. In the Phase II design, patients received no chemotherapy for at least two weeks and no immune or radiation therapy for at least four weeks prior to enrollment. Infusion pretreatment includes antihistamines, acetaminophen and anti-emetics, but no corticosteroid. Results: Six patients (F1,2,4–7) have been treated to date. F1 experienced CA-125 stabilization and approximately 20% reduction in her pelvic mass after two cycles, but was then withdrawn for hypoalbuminemia. F2 experienced CA-125 stabilization after two cycles and remains stable through four cycles. F4 experienced a 20% drop in CA-125 after two cycles, but then progressed. F5 progressed after one cycle. Treatment was recently initiated for F6–7. Existing immune data to date confirm that denileukin diftitox treatment in the Phase II trial is associated with reductions in phenotypic CD4+CD25hi blood Tregs and increased CD3+IFN-γ+ T cells consistent with our Phase I results. Studies of Treg function, T cell cytokine production and CD4+CD25+ T cell FOXP3 content using saved clinical material will be undertaken when full laboratory operations resume. The trial continues to accrue patients. Conclusions: These data suggest that denileukin diftitox depletes Tregs in ovarian cancer. The link between Treg depletion and any observed immune changes or clinical effects is under active investigation, and remains to be determined.

No significant financial relationships to disclose.
Impaired Ca++ mobilization in rituximab-resistant cells (RRCL) is associated with changes in the structure of CD20 antigen, down-regulation of Bax/Bak pro-apoptotic proteins and up-regulation of the endoplasmic reticulum (ER) Ca++ pump protein SERCA-3
Authors: Hernandez-Ilizaliturri F. J. , Kaur H. , Bhinder A. , Olejniczak S. , Knight J. , Czuczman M. S.
Published: March 2020

2516

Ca++ mobilization leading to apoptosis has been observed following rituximab biding to CD20 in lymphoma cells. Extending rituximab (R) exposure (e.g. as via R maintenance regimens) could potentially accelerate the emergence of resistance to rituximab. To define the molecular basis for rituximab resistance we developed several RRCL and demonstrated changes in CD20 expression/structure and membrane reorganization following rituximab therapy. Specifically, changes in the N-terminal and the C-terminal region of the internal domain of CD20 were observed in RRCL as determined by Western blotting using various antibodies recognizing epitopes located in the internal (GST77 and 1439) and external domain (B1) of CD20. In our current work we evaluated the effects that CD20 structure/expression changes have upon Ca++ mobilization. Extracellular and intracellular Ca++ mobilization was measured by flow cytometric analysis using FLUO-3 AM (acetoxymethyl ester). Optimization of the Ca++ indicator and calibration curves were performed for each cell line. Raji and RRCL were labeled under optimal conditions with FLUO-3 AM/Pluronic Acid F-127. Subsequently, cells were then re-suspended in HANKS media with or without Ca++ and exposed to rituximab or isotype (10μg/ml) ± human serum (25%). Surface CD20 expression was similar between Raji cells and RRCL. Expression of pro- or anti-apoptotic proteins and Ca++ regulatory proteins SERCA3, SERCA2 and Calreticulin was also determined by western blotting. In vitro exposure of Raji cells to rituximab + HS resulted in Ca++ mobilization even in Ca++ depleted media. Notably, Ca++ mobilization was impaired in RRCL when compared to Raji parental cells. In addition, down-regulation of Bax/Bak and up-regulation of SERCA3 was demonstrated in RRCL. Our data suggest that the acquirement of rituximab resistance is associated with changes in the intracellular domain of CD20 and in Ca++ regulator/pro-apoptotic proteins (SERCA3 and Bax/Bak) resulting in a decrease in the intracellular mobilization of Ca++.

No significant financial relationships to disclose.