Harald Enzmann

Analysis of signaling pathways related to cell proliferation stimulated by insulin analogs in human mammary epithelial cell lines

Insulin and insulin analogs stimulate proliferation of human mammary epithelial cells. We identified and analyzed the signaling pathways related to cell proliferation induced by regular insulin and by four insulin analogs presently approved for therapeutical use. Benign and malignant mammary cell lines showing different insulin receptor (IR) and IGF-I receptor (IGF-IR) expression patterns were studied. Cell proliferation was studied by crystal violet staining (BrdU-FACS analysis). Activation of insulin and IGF signaling pathways was studied by analysis of the phosphorylation status of IGF-IR and of key signaling proteins of the phosphoinositide 3-kinase (PI3K)/Akt and MAP kinase pathways, by the use of specific PI3K and MAP kinase inhibitors, and by silencing of IR and IGF-IR. Lantus stimulated the growth of MCF7 cells, which show high IGF-IR/IR ratio, significantly at 0.3 nmol/l, while regular insulin (Actrapid and bovine insulin) and other insulin analogs (Novorapid, Humalog, and Levemir) stimulated cell growth at 1.5-15 nmol/l concentrations. No difference between Lantus and the other insulin analogs was observed regarding growth stimulation of MCF10A cells showing low IGF-IR/IR ratio. Growth stimulation of MCF7 cells by Lantus was mainly due to strong activation of the IGF-IR and the MAP kinase pathway. Regular insulin and other insulin analogs tested activated mainly the IR and the PI3K/Akt pathway. We conclude that unlike regular insulin and other insulin analogs, Lantus strongly activates the IGF-IR and the MAP kinase pathway in MCF7 cells and is a strong mitogen for cells characterized by a high-IGF-IR/IR ratio.

Proliferative effects of insulin analogues on mammary epithelial cells.

Abstract Structural modification of insulin results in the generation of insulin analogues that show altered binding affinities to the insulin receptor and/or IGF-I receptor. As a consequence these insulin analogues may have increased mitogenic potency. Nine benign or malignant human mammary epithelial cells, which show different insulin receptor and IGF-I receptor expression patterns, were studied regarding mitogenicity of insulin and insulin analogues. Only insulin glargine showed a significantly higher proliferative effect on MCF-7 breast cancer cells compared to regular insulin among a panel of short- or long-acting insulin analogues, that are in clinical use.

Proliferative effect of Apidra (insulin glulisine), a rapid-acting insulin analogue on mammary epithelial cells.

The structural modification of insulin results in the generation of insulin analogues that show altered binding affinities to the insulin receptor and/or the IGF-I receptor, and as a consequence insulin analogues may have altered mitogenic potency. We analysed the proliferative effect of the rapid-acting insulin analogue Apidra® (insulin glulisine) on mammary epithelial cells. We show that Apidra and Actrapid® (recombinant human insulin) have similar proliferative effects on benign MCF10A and tumorigenic MCF7 cells and on epithelial cells of mouse mammary gland. Whereas Apidra and Actrapid induced similar activation of Erk1/2, activation of Akt/PKB by Apidra was significantly weaker compared to regular insulin. As AKT/PKB, an effector of the phosphoinositide 3-kinase pathway, mediates metabolic effects of insulin, we studied induction of hexokinase-2 in MCF7 cells and hexokinase-2 and hexokinase-4 in HepG2 cells by Actrapid and Apidra. Both genes were not significantly induced by Actrapid and Apidra in these cell lines.

Giving Patients’ Preferences a Voice in Medical Treatment Life Cycle: The PREFER Public–Private Project

Giving Patients’ Preferences a Voice in Medical Treatment Life Cycle : The PREFER Public–Private Project

Actual developments in European regulatory and health technology assessment of new cancer drugs: what does this mean for oncology in Europe?

ABSTRACT In the EU, the centralized procedure (CP) of the EMA is mandatory for marketing authorization for innovative anticancer drugs. However, numerous independent healthcare systems are in operation across the EU, and each HTA body follows its own methodologies and scientific value judgements in the assessment of the added value of an innovative anticancer drug. Initiatives to improve the interface between the different stakeholders are currently on the way, but are unlikely sufficient enough to overcome these fundamental problems.

Access to innovative oncology medicines in Europe

The incidence of cancer will increase within the next 5 years up to 15 million patients per year worldwide with a further increasing tendency. Therefore, there is a requirement for more emphasis on cancer prevention, research and effective anticancer drugs including a rapid licensing and market availability for the patients. In the EU, the centralized procedure (CP) of the European Medicine Agency (EMA) is mandatory for marketing authorization for anticancer drugs. A CP will result in one marketing authorization for all Member States granted by the European Commission. At variance, numerous independent healthcare sys-tems are in operation across the EU, and each Health Technology Assessment (HTA) body follows its own methodologies and scientific value judgements in the assessment of the ad-ditional clinical benefit of a new anticancer drug. Payer organisations in the various member states consider these assessments to a varying degree as input in their pricing and reim-bursement negotiations. At the same time international reference pricing and parallel trade have an inherent tendency to establish rather uniform price levels across member states. Consequently, drug access for patients differs considerably within the EU. Initiatives to im-prove the interface between the different stakeholders are currently on the way, but are unlikely sufficient to overcome these fundamental problems.

Differences in tissue distribution of iron from various clinically used intravenous iron complexes in fetal avian heart and liver.

Nanomedicines are more complex than most pharmacologically active substances or medicines and have been considered as non-biological complex drugs. For nanomedicines pivotal pharmacokinetic properties cannot be assessed by plasma concentration data from standard bioequivalence studies. Using intravenous iron complexes (IICs) as model we show that fetal avian tissues can be used to study time dependent tissue concentrations in heart and liver. Clear differences were found between equimolar doses of sucrose, gluconate or carboxymaltose coated iron particles. The range in tissue iron concentrations observed with these clinically widely used IICs provides an orientation as to what should be acceptable for any new IICs. Moreover, sensitivity of the experimental model was high enough to detect a 20% difference in tissue iron concentration. For the authorization of generic products under Article 10 (1) of Directive 2001/83/EC a plasma concentration of an active substance in the range of 80%-125% versus the reference product is usually considered acceptable. Based on its high discriminatory sensitivity this method was used to support a positive marketing authorization decision for a generic nanomedicine product.

Promoting Safe Early Clinical Research of Novel Drug Candidates: A European Union Regulatory Perspective

The European Medicines Agency (EMA) revises its guideline on minimizing risk in first‐in‐human trials to reflect changing practice and in light of a recent tragic incident.

Proceedings of the 2016 China Cancer Immunotherapy Workshop

Table of contentsA1 Proceedings of 2016 China Cancer Immunotherapy Workshop, Beijing, ChinaBin Xue, Jiaqi Xu, Wenru Song, Zhimin Yang, Ke Liu, Zihai LiA2 Set the stage: fundamental immunology in forty minutesZihai LiA3 What have we learnt from the anti-PD-1/PD-L1 therapy of advanced human cancer?Lieping ChenA4 Immune checkpoint inhibitors in lung cancerEdward B. GaronA5 Mechanisms of response and resistance to checkpoint inhibitors in melanomaSiwen Hu-LieskovanA6 Checkpoint inhibitor immunotherapy in lymphoid malignanciesWei DingA7 Translational research to improve the efficacy of immunotherapy in genitourinary malignanciesChong-Xian PanA8 Immune checkpoint inhibitors in gastrointestinal malignanciesWeijing SunA9 What’s next beyond PD-1/PDL1?Yong-Jun LiuA10 Cancer vaccines: new insights into the oldest immunotherapy strategyLei ZhengA11 Bispecific antibodies for cancer immunotherapyDelong LiuA12 Updates on CAR-T immunotherapyMichel SadelainA13 Adoptive T cell therapy: personalizing cancer treatmentCassian YeeA14 Immune targets and neoantigens for cancer immunotherapyRongfu WangA15 Phase I/IIa trial of chimeric antigen receptor modified T cells against CD133 in patients with advanced and metastatic solid tumorsMeixia Chen, Yao Wang, Zhiqiang Wu, Hanren Dai, Can Luo, Yang Liu, Chuan Tong, Yelei Guo, Qingming Yang, Weidong HanA16 Cancer immunotherapy biomarkers: progress and issuesLisa H. ButterfieldA17 Shaping of immunotherapy response by cancer genomesTimothy A. ChanA18 Unique development consideration for cancer immunotherapyWenru SongA19 Immunotherapy combinationRuirong YuanA20 Immunotherapy combination with radiotherapyBo LuA21 Cancer immunotherapy: past, present and futureKe LiuA22 Breakthrough therapy designation drug development and approvalMax NingA23 Current European regulation of innovative oncology medicines: opportunities for immunotherapyHarald Enzmann, Heinz Zwierzina