Tumor‐infiltrating lymphocytes for the treatment of metastatic cancer

Over the past few years melanoma incidence has been rising steadily, resulting in an increase in melanoma related mortality. Until recently, therapeutic options for metastatic melanoma were scarce. Chemotherapy and, in some countries, IL‐2 were the only registered treatment modalities. In the last five years, treatment with immunotherapy (anti CTLA‐4, anti PD‐1, or the combination of these antibodies) has shown very promising results and was able to improve survival in patients with metastatic melanoma. Adoptive cell therapy using tumor‐infiltrating lymphocytes is yet another, but highly promising, immunotherapeutic strategy for patients with metastatic melanoma. This review will discuss the development of TIL as a treatment option for melanoma, its mode of action and simplification over time, and the possibilities to expand this therapy to other types of cancer. Also, the future directions of TIL based therapies will be highlighted.     

Allo‐reactive T cells for the treatment of hematological malignancies

Several mechanisms can be responsible for control of hematological tumors by allo‐reactive T cells. Following allogeneic stem cell transplantation (alloSCT) donor T cells recognizing genetic disparities presented on recipient cells and not on donor cells are main effectors of tumor control, but also of the detrimental graft versus host disease (GVHD). Since after transplantation normal hematopoiesis is of donor origin, any T cell response directed against polymorphic antigens expressed on hematopoietic recipient cells but not on donor cells will result in an anti‐tumor response not affecting normal hematopoiesis. After fully HLA‐matched alloSCT, T cells recognizing polymorphic peptides derived from proteins encoded by genes selectively expressed in hematopoietic lineages may result in anti‐tumor responses without GVHD. Due to the high susceptibility of hematopoietic cells for T cell recognition, a low amplitude of the overall T cell response may also be in favor of the anti‐tumor reactivity in hematological malignancies. A mismatch between donor and patient for specific HLA‐alleles can also be exploited to induce a selective T cell response against patient (malignant) hematopoietic cells. If restricting HLA class II molecules are selectively expressed on hematopoietic cells under non‐inflammatory circumstances, allo HLA class‐II responses may control the tumor with limited risk of GVHD. Alternatively, T cells recognizing hematopoiesis‐restricted antigens presented in the context of mismatched HLA alleles may be used to treat patients with hematological cancers. This review discusses various ways to manipulate the allo‐immune response aiming to exploit the powerful ability of allo‐reactive T‐cells to control the malignancies without causing severe damage to non‐hematopoietic tissues.     

Volitinib,a potent and highly selective c‐Met inhibitor,effectively blocks c‐Met signaling and growth in c‐MET amplified gastric cancer patient‐derived tumor xenograft models

Purpose

To investigate the incidence of cMET gene copy number changes and protein overexpression in Chinese gastric cancer (GC) and to preclinically test the hypothesis that the novel, potent and selective cMET small‐molecule inhibitor volitinib, will deliver potent anti‐tumor activity in cMET‐dysregulated GC patient‐derived tumor xenograft (PDX) models.

Experimental design

A range of assays were used and included; in vitro cell line panel screening and pharmacodynamic (PD) analysis, cMET fluorescence in‐situ hybridization (FISH) and immunohistochemical (IHC) tissue microarray (TMA) analysis of Chinese GC (n = 170), and anti‐tumor efficacy testing and PD analysis of gastric PDX models using volitinib.

Results

The incidence of cMET gene amplification and protein overexpression within Chinese patient GC tumors was 6% and 13%, respectively. Volitinib displayed a highly selective profile across a gastric cell line panel, potently inhibiting cell growth only in those lines with dysregulated cMET (EC50 values 0.6 nM/L–12.5 nM/L). Volitinib treatment led to pharmacodynamic modulation of cMET signaling and potent tumor stasis in 3/3 cMET‐dysregulated GC PDX models, but had negligible activity in a GC control model.

Conclusions

This study provides an assessment of tumor cMET gene copy number changes and protein overexpression incidence in a cohort of Chinese GC patients. To our knowledge, this is the first study to demonstrate anti‐tumor efficacy in a panel of cMET‐dysregulated gastric cancer PDX models, using a novel selective cMET‐inhibitor (volitinib). Thus, the translational science presented here provides strong rationale for the investigation of volitinib as a therapeutic option for patients with GC tumors harboring amplified cMET.     

Cytokine‐regulated urokinase‐type‐plasminogen‐activator (uPA) production by human breast fibroblasts in vitro

It has been shown that, in breast stroma, urokinasetype plasminogen activator (uPA) mRNA is predominantly expressed by myofibroblasts located at the invasive areas of the tumor. To examine which factors present in a tumor environment are candidates responsible for the induction of these uPAproducing myofibroblasts, we studied in vitro the capacity of a paired panel of normal and tumorderived human breast fibroblasts to produce uPA protein and the myofibroblast marker smoothmuscleactin (SMA) in response to various cytokines implicated in the process of tissueremodeling during malignant transformation.We found that fibroblasts produced increased amounts of uPA protein after exposure to aFGF, bFGF, EGF, PDGFBB, and IFN, were unaffected in this respect by IL6, MCSF, GMCSF and Oncostatin M, and produced decreased amounts of uPA protein after exposure to IL1, TNF, IGFI, and IGFII. None of these cytokines were able to induce a striking increase in the fraction of SMApositive fibroblasts. On the other hand, 25pM TGF₁ increased the fraction of SMApositive fibroblasts 5fold in both normal and tumortissuederived fibroblasts. Nonetheless, the normalderived fibroblasts were unaffected in their uPAproducing capacity by TGF₁, and the tumorderived fibroblasts produced decreased amounts of uPA protein after exposure to this cytokine, implying that at least in vitro the myofibroblast phenotype is not a prerequisite for the production of uPA by human breast fibroblasts. In addition, we established that the basaluPAproduction of both normal and tumorderived fibroblasts was increased by autocrinely produced bFGFlike activity, and that the basaluPAproduction of at least the normalderived fibroblasts was decreased by autocrinely produced IGFlike activity.Altogether, our data suggest an active role for fibroblasts in the process of uPAdirected breast tumor proteolysis.     

Glycan‐related gene expression signatures in breast cancer subtypes; relation to survival

Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes – and look for their association to clinical parameters.Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non‐malignant breast tissue samples were analysed. Results were validated in 1960 breast carcinomas. 419 genes encoding glycosylation‐related proteins were selected.The DCIS samples appeared expression‐wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N‐glycans when compared to non‐malignant samples. In‐situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. Subtype‐specific expression patterns revealed down‐regulation of genes encoding glycan‐binding proteins in the luminal A and B subtypes. Clustering basal‐like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors – as demonstrated by association of B3GNT5 and UGCG genes to patient survival.In conclusion, most glycan‐specific changes occur early in the carcinogenic process. We have identified glycan‐related alterations specific to breast cancer subtypes including a prognostic signature for two basal‐like subgroups. Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients.     

Incremental cost‐effectiveness of double‐reading mammograms

Background. Double reading is a widely used criterion standard in breast cancer screening despite a lack of evidence of the costeffectiveness of the second reading. This study evaluates the incremental costeffectiveness of such a strategy.Design. Costeffectiveness analysis: Nationwide populationbased semiannual screening program for women aged 50–59 in Finland. Participation rate was 91%. All mammograms (95,423) performed during 1990–1995 in three screening centers of the Finnish Cancer Society were read by two radiologists with gradings recorded. The effectiveness of the double reading was the difference in cancers detected in the double compared to that of the single reading. Incremental costs of the double reading for the health care and nonhealth care and the time costs were estimated. The main outcome measure was the incremental cost per additional cancer found as a result of the doublereading strategy.Results. The total number of cancers detected with the double and single reading were 290 and 261, respectively. A significantly higher ratio of carcinoma in situ was the causative pathology in cancers detected only by the second reader. The cost per cancer detected with a single reading was US$ 18,340. The incremental cost of any additional cancer found was US$ 25,523, that is, a 39% higher cost per additional cancer found by double reading.Conclusions. The additional cost per cancer detected by double reading is not drastically higher than with single reading. However, the additional cost per life year saved may be much higher.     

Liposome-coated lipoplex–based carrier for antisense oligonucleotides

The chemical nature of genetic drugs (e.g. antisense oligonucleotides, siRNA, vectors) requires a suitable carrier system to protect them from enzymatic degradation without changing their properties and enable efficient delivery into target cells. Lipid vectors for nucleic acid delivery that have been widely investigated for years can be very effective. As the majority of attempts made in the field of cancer gene therapy have focused on solid tumors, while blood cancer cells have attracted less attention, the latter became the subject of our investigation. The lipid carrier proposed here is based on liposomes constructed by others but the lipid composition is original. A liposome-coated lipoplex (L-cL) consists of a core arising from complexation of positively charged lipid and negatively charged oligodeoxynucleotide (ODN) or plasmid DNA coated by a neutral or anionic lipid bilayer. Moreover, our lipid vector demonstrates size stability and is able to retain a high content of enclosed plasmid DNA or antisense oligodeoxynucleotides (asODNs). Observed transfection efficacies of the tested preparation using a plasmid coding for fluorescent protein were up to 60-85% of examined leukemia cells (Jurkat T and HL-60 lines) in the absence or the presence of serum. When BCL‑2 asODN was encapsulated in the L-cL, specific silencing of this gene product at both the mRNA and protein level and also a markedly decreased cell survival rate were observed in vitro. Moreover, biodistribution analysis in mice indicates prolonged circulation characteristic for PEG-modified liposomal carriers. Experiments on tumor-engrafted animals indicate substantial inhibition of tumor growth.     

MTA1 regulates higher‐order chromatin structure and histone H1‐chromatin interaction in‐vivo

In the current study, for the first time, we found that metastasis‐associated gene 1 (MTA1) was a higher‐order chromatin structure organizer that decondenses the interphase chromatin and mitotic chromosomes. MTA1 interacts dynamically with nucleosomes during the cell cycle progression, prominently contributing to the mitotic chromatin/chromosome structure transitions at both prophase and telophase. We showed that the decondensation of interphase chromatin by MTA1 was independent of Mi‐2 chromatin remodeling activity. H1 was reported to stabilize the compact higher‐order chromatin structure through its interaction with DNA. Our data showed that MTA1 caused a reduced H1‐chromatin interaction in‐vivo. Moreover, the dynamic MTA1‐chromatin interaction in the cell cycle contributed to the periodical H1‐chromatin interaction, which in turn modulated chromatin/chromosome transitions. Although MTA1 drove a global decondensation of chromatin structure, it changed the expression of only a small proportion of genes. After MTA1 overexpression, the up‐regulated genes were distributed in clusters along with down‐regulated genes on chromosomes at parallel frequencies.     

Subtype‐dependent prognostic relevance of an interferon‐induced pathway metagene in node‐negative breast cancer

The majority of gene expression signatures developed to predict the likelihood to relapse in breast cancer (BC) patients assigns a high risk score to patients with Estrogen Receptor (ER) negative or highly proliferating tumors. We aimed to identify a signature of differentially expressed (DE) metagenes, rather than single DE genes, associated with distant metastases beyond classical risk factors.We used 105 gene expression profiles from consecutive BCs to identify metagenes whose prognostic role was defined on an independent series of 92 ESR1+/ERBB2− node‐negative BCs (42 cases developing metastases within 5 years from diagnosis and 50 cases metastasis‐free for more than 5 years, comparable for age, tumor size, ER status and surgery). Findings were validated on publicly available datasets of 684 node‐negative BCs including all the subtypes.Only a metagene containing interferon‐induced genes (IFN metagene) proved to be predictive of distant metastasis in our series of patients with ESR1+/ERBB2− tumors (P = 0.029), and such a finding was validated on 457 ESR1+/ERBB2− BCs from public datasets (P = 0.0424). Conversely, the IFN metagene was associated with a low risk of metastasis in 104 ERBB2+ tumors (P = 0.0099) whereas it did not prove to significantly affect prognosis in 123 ESR1−/ERBB2− tumors (P = 0.2235). A complex prognostic interaction was revealed in ESR1+/ERBB2− and ERBB2+ tumors when the association between the IFN metagene and a T‐cell metagene was considered.The study confirms the importance of analyzing prognostic variables separately within BC subtypes, highlights the advantages of using metagenes rather than genes, and finally identifies in node‐negative ESR1+/ERBB2− BCs, the unfavorable role of high IFN metagene expression.     

Inhibition of PARP1‐dependent end‐joining contributes to Olaparib‐mediated radiosensitization in tumor cells

Poly‐ADP‐ribose‐polymerase inhibitors (PARPi) are considered to be optimal tools for specifically enhancing radiosensitivity. This effect has been shown to be replication‐dependent and more profound in HR‐deficient tumors. Here, we present a new mode of PARPi‐mediated radiosensitization which was observed in four out of six HR‐proficient tumor cell lines (responders) investigated, but not in normal cells. This effect is replication‐independent, as the radiosensitization remained unaffected following the inhibition of replication using aphidicolin. We showed that responders are radiosensitized by Olaparib because their DSB‐repair is switched to PARP1‐dependent end‐joining (PARP1‐EJ), as evident by (i) the significant increase in the number of residual γH2AX foci following irradiation with 3Gy and treatment with Olaparib, (ii) the enhanced enrichment of PARP1 at the chromatin after 3Gy and (iii) the inhibition of end‐joining activity measured by a specific reporter substrate upon Olaparib treatment. This is the first study which directly demonstrates the switch to PARP1‐EJ in tumor cells and its contribution to the response to Olaparib as a radiosensitizer, findings which could widen the scope of application of PARPi in tumor therapy.     

Metastatic bone pain palliation with 89‐Sr and 186‐Re‐HEDP in breast cancer patients

Aim. The study evaluates the therapeutic efficacy of Strontium89chloride (89Sr) and 186Re1,1hydroxyethylidene diphosphonate (186ReHEDP) in the palliation of painful bone metastases from breast cancer. Patients and methods. Fifty patients with painful multifocal bone metastases from breast cancer entered the study and were randomized into two groups according to the radiopharmaceutical used: 148 MBq 89Sr i.v. (Group A: 25 patients) and 1406 MBq 186ReHEDP i.v. (Group B: 25 patients). Pain palliation was evaluated on the basis of the Wisconsin pain test improvement at two months and response was graded as complete, partial, minimal or absent. Hematological toxicity and side effects were reported according to WHO guidelines. Results. The global response rate was 84% (21/25) for 89Sr and 92% (23/25) for 186ReHEDP, respectively. The onset of pain palliation appeared significantly earlier in Group B (p<0.0001). The duration of pain relief ranged from two months to 14 months (mean of 125 days with a median value of 120 days) in Group A and from one month to 12 months (mean of 107 days with a median value of 60 days) in Group B (p=0.39). A moderate hematological toxicity was apparent in both groups. Platelet and white blood cell counts returned to baseline levels within 12 weeks after 89Sr administration and 6 weeks after 186ReHEDP administration (p<0.01). Conclusions. Both 89Sr and 186ReHEDP are effective and safe in bone pain palliation in breast cancer with the latter showing a significantly faster onset of pain relief.     

“脑型”肺癌的诊断问题(附10例临床分析)

 脑转移癌并不少见,其发生率约占脑瘤的13.5～37.5%,占恶性肿瘤尸检资料的5～10%。有时原发肿瘤病灶隐匿,以脑转移损害作为首发症状,常被误认为原发性脑瘤而手术,诊断每感困难。国内外资料证明脑转移癌以肺癌占首位,肺癌以神经精神症状而首先发病者并不少见,我们一年中就收集到10例病人,多误诊为其它神经精神疾病。     

伴rob(13;22)(q10;q10)的治疗相关急性髓系白血病1例并文献复习

目的提高对治疗相关急性髓系白血病(AML)的认识。方法回顾性分析烟台毓璜顶医院收治的1例横纹肌肉瘤治疗后继发AML伴染色体核型rob(13;22)(q10;q10)患儿的临床资料,并进行文献复习。结果该例女性患儿6岁5个月,在2018年1月诊断为横纹肌肉瘤,接受规范化疗30个月后确诊为治疗相关AML,染色体核型45,XX,t(11;19)(q23;q13),rob(13;22)(q10;q10),融合基因MLL⁃ELL阳性。接受DA(阿糖胞苷+柔红霉素)方案化疗后无明显改善,家属放弃治疗,患儿死亡。结论治疗相关AML多与应用鬼臼毒素类(依托泊苷等)药物化疗、联合放疗等因素有关。具有高危细胞遗传学因素的治疗相关AML预后差。     

5‐Fluorouracil preferentially sensitizes mutant KRAS non‐small cell lung carcinoma cells to TRAIL‐induced apoptosis

Mutations in the KRAS gene are very common in non–small cell lung cancer (NSCLC), but effective therapies targeting KRAS have yet to be developed. Interest in tumor necrosis factor–related apoptosis‐inducing ligand (TRAIL), a potent inducer of cell death, has increased following the observation that TRAIL can selectively kill a wide variety of human cancer cells without killing normal cells both in vitro and in xenograft models. However, results from clinical trials of TRAIL‐based therapy are disappointingly modest at best and many have demonstrated a lack of therapeutic benefit. Current research has focused on selecting a subpopulation of cancer patients who may benefit from TRAIL‐based therapy and identifying best drugs to work with TRAIL. In the current study, we found that NSCLC cells with a KRAS mutation were highly sensitive to treatment with TRAIL and 5‐fluorouracil (5FU). Compared with other chemotherapeutic agents, 5FU displayed the highest synergy with TRAIL in inducing apoptosis in mutant KRAS NSCLC cells. We also found that, on a mechanistic level, 5FU preferentially repressed survivin expression and induced expression of TRAIL death receptor 5 to sensitize NSCLC cells to TRAIL. The combination of low‐dose 5FU and TRAIL strongly inhibited xenograft tumor growth in mice. Our results suggest that the combination of TRAIL and 5FU may be beneficial for patients with mutant KRAS NSCLC.     

Novel circulating microRNA signature as a potential non‐invasive multi‐marker test in ER‐positive early‐stage breast cancer: A case control study

Introduction

There are currently no highly sensitive and specific minimally invasive biomarkers for detection of early‐stage breast cancer. MicroRNAs (miRNAs) are present in the circulation and may be unique biomarkers for early diagnosis of human cancers. The aim of this study was to investigate the differential expression of miRNAs in the serum of breast cancer patients and healthy controls.

Methods

Global miRNA analysis was performed on serum from 48 patients with ER‐positive early‐stage breast cancer obtained at diagnosis (24 lymph node‐positive and 24 lymph node‐negative) and 24 age‐matched healthy controls using LNA‐based quantitative real‐time PCR (qRT‐PCR). A signature of miRNAs was subsequently validated in an independent set of 111 serum samples from 60 patients with early‐stage breast cancer and 51 healthy controls and further tested for reproducibility in 3 independent data sets from the GEO Database.

Results

A multivariable signature consisting of 9 miRNAs (miR‐15a, miR‐18a, miR‐107, miR‐133a, miR‐139‐5p, miR‐143, miR‐145, miR‐365, miR‐425) was identified that provided considerable discrimination between breast cancer patients and healthy controls. Further, the ability of the 9 miRNA signature to stratify samples from breast cancer patients and healthy controls was confirmed in the validation set (p = 0.012) with a corresponding AUC = 0.665 in the ROC‐curve analysis. No association between miRNA expression and tumor grade, tumor size, menopausal‐ or lymph node status was observed. The signature was also successfully validated in a previously published independent data set of circulating miRNAs in early‐stage breast cancer (p = 0.024).

Conclusions

We present herein a 9 miRNA signature capable of discriminating between ER‐positive breast cancer and healthy controls. Using a specific algorithm based on the 9 miRNA signature, the risk for future individuals can be predicted. Since microRNAs are highly stable in blood components, this signature might be useful in the development of a blood‐based multi‐marker test to improve early detection of breast cancer. Such a test could potentially be used as a screening tool to identify individuals who would benefit from further diagnostic assessment.     

Successful co‐treatment with LHRH‐agonist for ovarian over‐stimulation and cystic formation in premenopausal tamoxifen exposure

The present study evaluates the potential beneficial effect of cotreatment with LHRHagonist in resolving premenopausal tamoxifen's induced supraphysiological serum 17 estradiol levels and persistent ovarian cysts. Ultrasonographic and serum hormonal evaluations were performed before, during, and following three consecutive injections of long acting LHRHagonist administered to 14 premenopausal breast cancer patients treated with tamoxifen, who had supraphysiological serum 17 estradiol levels and simultaneous persistent ovarian cysts. Within 3 weeks of the first LHRHagonist injection, all patients had menopausal serum estradiol levels. Ovarian cysts completely disappeared within 2 months following the first injection. Following the discontinuation of LHRHagonist cotreatment, serum estradiol levels remained in physiological levels and the ovaries remained a normal size in 64.3% of the patients for 13.3 ± 11.5 months. 28.6% of the patients had a gradual reappearance of high serum estradiol levels and of ovarian cysts, and were, therefore, treated with a second course of LHRHagonist. Following the second course, serum estradiol levels remained in physiological levels and the ovaries remained a normal size for 8–15 months. It is concluded that short duration of cotreatment with long acting LHRHagonist administered to premenopausal breast cancer patients treated with tamoxifen, successfully resolved the tamoxifeninduced supraphysiological serum 17 estradiol levels and the ovarian cysts.affiliated with Sackler Faculty of Medicine, Tel Aviv University