Cytotoxic Chemotherapeutic Agents Research Articles

Drug Release Studies of SC‐514 PLGA Nanoparticles

Background A major problem associated with prostate cancer treatment is drug resistance leading to prostate cancer metastasis. Prostate cancer-targeted drug delivery system is one way to address this problem. Traditional drug delivery systems face many challenges, such as inability to control the release rate, inaccurate targeting, susceptibility to the microenvironment, and poor stability of the drug. Hence, there is a need to formulate a drug delivery system and functionalize the surface of delivery system for a better control of drug release. Targeted drug delivery of nanoparticles decorated with site-specific recognition ligands is of considerable interest to minimize cytotoxicity of chemotherapeutics in the normal cells. This study was designed to measure drug release of SC-514-PLGA nanoparticles and conjugate SC-514 polylactic-co-glycolic acid (PLGA) nanoparticles loaded with NF-KB antibody and fats. This study explored new methods to quantify the SC-514 drug released from SC-514-PLGA nanoparticles. The anti-proliferative effects of the SC-514 drug released by the nanoparticle treatments were investigated in PC-3 cells and cord blood cells. MDR expression in PC-3 cells was investigated after treatment with free SC-514 and SC-514 from SC-514-PLGA nanoparticles. Methods SC-514drug release was studied under in vitro conditions in phosphate buffer solution (PBS) at a pH of 7.4, using dialysis method. Liquid chromatography–mass spectrometry (LC-MS) was utilized as the standard method to quantify SC-514 drug released. Furthermore, other experimental methods utilized to quantify SC-514 drug released included: colony assay, wound healing assay, and transwell migration and invasion assays. MTT calorimetric assay was utilized to investigate the impact of the nanoparticle formulations (SC-514-PLGA, SC-514-PLGA-NF-KB, SC-514-PLGA-Fat) on PC-3 cells and cord blood cells. In addition, in vitro cellular uptake of fluorescent nanoparticle formulations in PC-3 and cord blood cells were examined with confocal fluorescence microscope. Results The ligand conjugated nanoparticles showed considerable reduction of tumor growth and toxicity (side-effect of the drug treatment in prostate cancer) of SC-514 drug in prostate cancer treatment. Our results demonstrated that prepared drug delivery system possesses a much lower toxicity, better prostate cancer controlled-release behaviors and confers higher solubility to SC-514 than free SC-514. SC-514 released from SC-514-PLGA, PLGA-SC-514-NF-KB, and PLGA-SC-514-Fat significantly inhibited tumor growth when compared to free SC-514. A more significant reduction of cell viability was recorded in PC-3 cells by all treatments (Free SC-514, SC-514-PLGA, and SC-514-PLGA-NF-KB) compared to cord blood cells, except for SC-514-PLGA-Fat nanoparticle formulation.SC-514 drug from SC-514-PLGA nanoparticles reduced the expression of MDR proteins than free SC-514. Conclusion Encapsulation of SC-514 with PLGA polymer increased the anti-cancer therapeutic effects of SC-514 drug. Burst release from SC-514-PLGA nanoparticles can be controlled by functionalization. New methods of quantifying drug release maybe as effective as the standard methods of drug quantification, such as LC/MS method.

Cytotoxic chemotherapeutic agents and the EGFR-TKI osimertinib induce calreticulin exposure in non–small cell lung cancer

ObjectivesSynergistic anticancer efficacy of combination treatment with immune checkpoint inhibitors (ICIs) and platinum-based chemotherapy in patients with advanced non–small cell lung cancer (NSCLC) may be attributable in part to the phenomenon of immunogenic cell death (ICD), which is characterized by the release of damage-associated molecular patterns (DAMPs) from dying tumor cells. The ability of cytotoxic chemotherapeutic agents and molecularly targeted drugs such as epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) to induce DAMPs during the treatment of NSCLC has remained unclear, however. Materials and methodsWe investigated the ability of seven cytotoxic chemotherapeutic agents and the third-generation EGFR-TKI osimertinib to induce translocation of the DAMP calreticulin to the cell surface in multiple NSCLC cell lines. The plasma concentration of soluble CRT in advanced NSCLC patients treated with cytotoxic chemotherapy or osimertinib was measured. ResultsAntimetabolites and microtubule inhibitors induced expression of CRT at the cell surface (ecto-CRT) to a greater extent than did platinum agents in six NSCLC cell lines, exhibiting higher up-regulation of phosphorylation of eukaryotic initiation factor-2α (eIF2α). Ecto-CRT expression was positively correlated with apoptosis induction in NSCLC cells treated with these various chemotherapeutic agents. The drug-induced up-regulation of ecto-CRT in NSCLC cells was attenuated by the pan-caspase inhibitor Z-VAD-FMK. Osimertinib similarly increased ecto-CRT expression in association with apoptosis induction in five EGFR-mutated NSCLC cell lines. Furthermore, the plasma concentration of soluble CRT in 16 NSCLC patients treated with single-agent pemetrexed or docetaxel and in nine EGFR-mutated NSCLC patients treated with osimertinib was increased after treatment onset. ConclusionOur findings indicate that antimetabolites, microtubule inhibitors, and osimertinib are effective inducers both of CRT exposure in NSCLC cell lines and of soluble CRT release in patients with advanced NSCLC, suggesting that these agents might prove effective for promotion of antitumor immunity in combination immunotherapy.

Interim results of a multicenter retrospective-prospective observational post-marketing study of Extimia® BIOCAD (INN: empegfilgrastim) to evaluate safety and efficacy in patients with lymphoproliferative diseases receiving cytotoxic therapy

Aim. To assess the efficacy and safety of using the drug Extimia BIOCAD (international nonproprietary name INN: empegfilgrastim) in order to reduce the frequency and duration of neutropenia, the frequency of febrile neutropenia (FN) and infections manifested by FN in patients with lymphoproliferative diseases receiving myelosuppressive therapy.
 Materials and methods. This publication presents the interim results of a multicenter retrospective prospective observational post-marketing study of the safety and efficacy of the drug Extimia BIOCAD (INN: empegfilgrastim) in patients with lymphoproliferative diseases receiving cytotoxic therapy (LEGERITY). The interim data analysis included 40 patients with lymphoproliferative diseases (Hodgkins lymphoma, diffuse large B-cell lymphoma, multiple myeloma, primary mediastinal large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia, splenic marginal zone lymphoma), who were treated in ten research centers of the Russian Federation (Moscow, St. Petersburg, regional clinics). The median age of patients was 48 (2172) years, 13/40 (32.5%) patients belonged to the older age group (60 years). Patients had functional status on the ECOG scale of 02 and received at least 2 chemotherapy injections against the background of prophylaxis with empegfilgrastim. Empegfilgrastim was administered at a dose of 7.5 mg subcutaneously once 24 hours after the end of the administration of cytotoxic chemotherapeutic agents. Primary endpoint: frequency of neutropenia 35 degrees of severity; secondary endpoints: frequency of FN; frequency of severe infections (34 stages); frequency of antibiotic prescription; relative dose intensity of therapy of the conducted chemotherapy courses; the incidence of all adverse reactions in patients who received at least one dose of the study drug empegfilgrastim; the incidence of all serious adverse reactions in patients who received at least one dose of the study drug empegfilgrastim; the incidence of CTCAE 5.0 grade 34 HP in patients who received at least one dose of the study drug empegfilgrastim; discontinuation rate of study drug empegfilgrastim due to adverse reactions.
 Results. The results of this study demonstrate that the incidence of neutropenia of 3 degree of severity after the 1st cycle of chemotherapy developed in 2/40 patients (5%) and as a result of high-dose therapy (R-DHAP). Neutropenia of any severity was reported in 5/40 patients (12.5%). Cases of FN development have not been registered. Severe infections (mucositis, enteropathy, pneumonia, etc.), as well as the use of antibacterial and antifungal drugs during 1 cycle of chemotherapy and in the inter-course period after 1 cycle of therapy were not recorded in any patient. The next course of myelosuppressive therapy was not delayed due to the development of neutropenia in any of the patients during the study. Adverse events, according to the researcher, associated with the use of empegfilgrastim, were registered in 2/40 patients (5%): moderate generalized pain syndrome (diffuse pain) of 1 severity and in one case ossalgia of 2 severity. No serious adverse reactions were reported.
 Conclusion. The results of the interim analysis of the study demonstrate the high efficacy of the first Russian original pegylated granulocyte colony-stimulating factor empegfilgrastim after a single administration of a fixed dose in the treatment of patients with aggressive and indolent lymphomas. The drug has a favorable tolerance profile in any age group of patients, especially in elderly patients. Administration of empegfilgrastim as a prophylaxis of neutropenia in patients with lymphoproliferative diseases receiving myelosuppressive therapy of varying intensity can reduce the burden on medical personnel, improve patient adherence to treatment, and contribute to the implementation of the therapeutic plan.

Abstract GS1-07: Treatment persistence of residual breast tumors through an embryonic diapause-like cancer cell state with suppressed myc activity

Abstract Systemic breast cancer treatments often fail to achieve complete and sustained responses due to drug-persistent residual tumor foci, the “seed” for eventual relapse. Recent clinical studies have revealed that the chemo-persistent tumor cells undergo extensive transcriptional reprogramming in response to neo-adjuvant treatment; however, the impact of this acquired molecular signature on the ability of residual cancer cells to survive during therapy is not clear. To further study the molecular hallmarks of chemo-persistent cancer cells, we analyzed the transcriptional signatures of post-treatment residual tumors in a large number of breast cancer patients from several neoadjuvant clinical studies. We observed that the treatment-persistent tumor cells had a distinct cellular state which molecularly resembles that of the embryonic diapause, a dormant stage of transiently suspended development in undifferentiated epiblasts triggered by stress and induced by suppression of Myc activity and overall biosynthesis. Remarkably, the propensity for residual tumors with an embryonic diapause-like (EDL) molecular signature was significantly associated with worse outcome in breast cancer patients. To dissect this distinct cancer cell state, we developed novel in vitro models using 3D breast cancer organoids which responded to cytotoxic treatment by generating longitudinally-persistent residual organoid fractions that phenotypically and molecularly simulated the in vivo emergence of post-treatment residual tumors in preclinical and clinical settings. The treatment-persistent tumor fractions in cancer organoids and in the respective in vivo xenografts did not exhibit significant genetic changes compared to baseline tumor cells, but had reduced apoptotic priming and an EDL transcriptional reprograming similar to that of residual tumors in patients. Similarly to embryonic diapause, residual persistent fractions in cancer organoids, xenografts and patient tumors had markedly suppressed Myc transcriptional output and biosynthetic levels. Ectopic induction of MYC expression enhanced acute chemotherapeutic cytotoxicity in breast cancer organoids. Conversely, suppression of MYC or pharmacological inhibition of Myc transcriptional co-activators, BET bromodomains, abrogated chemotherapeutic cytotoxicity and induced in breast cancer cells an EDL molecular signature characterized by below-baseline redox stress levels which were maintained during drug exposure. High-throughput interrogation of residual persistent breast cancer organoids indicated broad refractoriness to specific anticancer drug classes thought to operate through induction of cellular stress (e.g. agents targeting DNA or DNA-repair). However, maintaining the residual cells in dormancy after completion of cytotoxic chemotherapy via inhibition of Brd4/Myc axis or pharmacologically interfering with the diapause-like transcriptional reprogramming of treatment-persistent cancer cells represent potential therapeutic strategies to target chemo-persistent tumor cells. Overall, our study shows that breast tumors dynamically co-opt the stress survival mechanism of embryonic diapause to persist during treatment, and reveals an unexpected role of Myc as regulator of cancer cell entry into transient drug-refractory dormancy. The diapause-like persister organoid cancer models provide ex vivo tractability for studying the otherwise elusive, dormant, drug-refractory residual tumors, with potential implications in personalized medicine and drug discovery. Citation Format: Eugen Dhimolea, Ricardo De Matos Simoes, Dhvanir Kansara, Juliette Bouyssou, Jennifer Roth, Michal Sheffer, Rinath Jeselsohn, Nathanael Gray, Ulrich Steidl, Boris Bartholdy, Myles Brown, Aedin Culhane, Constantine Mitsiades. Treatment persistence of residual breast tumors through an embryonic diapause-like cancer cell state with suppressed myc activity [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr GS1-07.

Repurposing approved drugs for cancer therapy.

BackgroundMany drugs approved for other indications can control the growth of tumor cells and limit adverse events (AE).Data sourcesLiterature searches with keywords ‘repurposing and cancer’ books, websites: https://clinicaltrials.gov/, for drug structures: https://pubchem.ncbi.nlm.nih.gov/Areas of agreementIntroducing approved drugs, such as those developed to treat diabetes (Metformin) or inflammation (Thalidomide), identified to have cytostatic activity, can enhance chemotherapy or even replace more cytotoxic drugs. Also, anti-inflammatory compounds, cytokines and inhibitors of proteolysis can be used to control the side effects of chemo- and immuno-therapies or as second-line treatments for tumors resistant to kinase inhibitors (KI). Drugs specifically developed for cancer therapy, such as interferons (IFN), the tyrosine KI abivertinib TKI (tyrosine kinase inhibitor) and interleukin-6 (IL-6) receptor inhibitors, may help control symptoms of Covid-19.Areas of controversyBetter knowledge of mechanisms of drug activities is essential for repurposing. Chemotherapies induce ER stress and enhance mutation rates and chromosome alterations, leading to resistance that cannot always be related to mutations in the target gene. Metformin, thalidomide and cytokines (IFN, tumor necrosis factor (TNF), interleukin-2 (IL-2) and others) have pleiomorphic activities, some of which can enhance tumorigenesis. The small and fragile patient pools available for clinical trials can cloud the data on the usefulness of cotreatments.Growing pointsBetter understanding of drug metabolism and mechanisms should aid in repurposing drugs for primary, adjuvant and adjunct treatments.Areas timely for developing researchOptimizing drug combinations, reducing cytotoxicity of chemotherapeutics and controlling associated inflammation.

Current Use of Total Body Irradiation in Haploidentical Allogeneic Hematopoietic Stem Cell Transplantation.

Total body irradiation (TBI) is included in the conditioning regimen for allogeneic hematopoietic stem cell transplantation (HSCT), with unique advantages such as uniform distribution over the whole body and decreased exposure to cytotoxic chemotherapeutic agents. For individuals who lack matched sibling or matched unrelated donors, the use of haploidentical donors has been increasing despite challenges such as graft rejection and graft-versus-host disease (GVHD). Although a limited number of studies have been performed to assess the clinical role of TBI in haploidentical HSCT, TBI-based conditioning showed comparable results in terms of survival outcomes, rate of relapse, and GVHD in diverse hematologic malignancies such as leukemia, lymphoma, and multiple myeloma. Advances in supportive care, along with recent technical improvements such as restriction of maximum tolerated dose, appropriate fractionation, and organ shielding, help to overcome diverse adverse events related to TBI. Post-transplantation cyclophosphamide was used in most studies to reduce the risk of GVHD. Additionally, it was found that post-transplantation rituximab may improve outcomes in TBI-based haploidentical HSCT, especially in patients with B-cell lymphoma. Along with the advances of techniques and strategies, the expansion of age restriction would be another important issue for TBI-based haploidentical HSCT considering the current tendency toward increasing age limitation and lack of matched donors. This review article summarizes the current use and future perspectives of TBI in haploidentical HSCT.

How to approach shared decision making when determining consolidation, maintenance therapy, and transplantation in acute myeloid leukemia.

Until recently, treatment options for patients with acute myeloid leukemia (AML) were limited to cytotoxic chemotherapeutic agents that possessed little specificity for the cytogenetic and molecular mutations known to risk stratify patients with this disease. With the approval of multiple new therapies, not only have the agents that we treat patients with changed, but the way we talk about these options, decide on, and manage therapy has also been transformed. Given these complexities, it is important that we help patients make an informed decision by weighing the risk of relapse with patient wishes and desired quality of life. Shared decision making (SDM) is an approach to medical decision making for those situations in which most clinicians would agree that there is more than 1 correct choice for a patient. Here we review the principles of SDM and provide an overview of the 3-talk model and how it may be incorporated into the care of patients with AML.

Matrine Inhibitory Effect on Self-renewal and Re-sensitization of 5-FU Resistant NSCLC Stem Cells were through Let-7b dependent Downregulation of CCND1

ABSTRACT Matrine is one of the major alkaloids extracted from Sophora flavescens Ait of the traditional Chinese medicine, was the main chemical ingredient of compounds of Kushen injection. The Matrine is considered as a promising therapeutic agent for curing nonsmall cell lung cancer (NSCLC), used either alone or combined with chemotherapeutic agents. In the present study, we focused on the possible roles of Matrine exerted on the self-renewal ability of stem-like cells of the NSCLC group, as well as the cytotoxicity of chemotherapeutic agents, in vitro and in vivo. Here we reported that Matrine inhibits cancer stem-like cell (CSC) properties through upregulation of Let-7b and suppression of the Wnt pathway. Overexpression of Let-7b suppressed the ability of tumorsphere formation, decreased Wnt pathway activation through inhibiting its transcriptional activity in lung CSCs. Further studies revealed that Let-7b directly targeted CCND1 and decreased its expression, whereas Matrine increased Let-7b levels and followed by inactivation of the CCND1/Wnt signaling pathway and inhibition of EMT, which was characterized by loss of epithelial markers and acquisition of a mesenchymal phenotype in lung CSCs. What is more, we found that Matrine increased Let-7b level in an endoribonuclease DICER1-dependent manner. And xenografts in nude mice evidenced that Matrine increased the sensitivity of lung CSCs to 5-FU and inhibited the accumulation of CCND1 in tumor tissues induced by 5-FU. Taken together, these data illustrate the role of Let-7b in regulating lung CSCs traits and DICER1/let-7/CCND1 axis in Matrine or in combination with 5-FU intervention of lung CSCs’ expansion, helping to fulfill the anti-cancer action of Matrine.

Abstract 6388: MERTK and ROCK inhibitors mediate synergistic AML cell death and enhance therapeutic activity in combination with cytotoxic chemotherapy

Abstract Survival rates are suboptimal for adult and pediatric patients with acute myeloid leukemia (AML). Cytotoxic chemotherapies have short and long-term side effects and are often contraindicated for elderly patients. Targeted agents can reduce toxicity compared to chemotherapy; however, resistance to single-agents often develops, thus combination therapies may provide more durable responses. To increase efficacy and reduce exposure to cytotoxic agents, a targeted combination therapy (MERTK/ROCKi) was utilized alone and with doxorubicin or etoposide, frontline cytotoxic chemotherapeutics routinely used for AML treatment. MERTK is aberrantly expressed in >80% of AML patient samples and MERTK inhibitors are in clinical development. Both MERTK and rho-associated, coiled-coil-containing protein kinases 1 and 2 (ROCK1/2) regulate actin/microtubule dynamics and cell cycle progression and shMERTK or siROCK1 knockdown induces apoptosis in AML cells. Here, analysis of The Cancer Genome Atlas database revealed poorer overall survival in patients with higher levels of ROCK1 mRNA (p<0.01, n=172). MERTK inhibitors (MRX-2843 or UNC3997) and ROCK1/2 inhibitors (RKI-1447 or GSK269962A) synergized to reduce cell density in 4 of 6 AML cell lines tested and had an additive effect in the remaining cell lines. Flow cytometric analysis of cells stained with popro-1-iodide and propidium iodide (PI) dyes revealed synergistic induction of cell death in KG-1 (p<0.05), OCI-AML5 (p<0.05), and NB4 (p<0.1) cell cultures and an additive effect in Kasumi-1 cultures. Cells treated with the combination therapy failed to expand, even when cultured in the absence of inhibitor(s), and cell density was reduced in response to treatment with the combination therapy compared to single agents (p<0.01). Flow cytometric analysis of DNA content revealed an increased fraction of cells with G2/M DNA content in cultures treated with MRX-2843 (p<0.01). This effect was more pronounced in cultures treated with the combination therapy (p<0.05-p<0.0001) and was accompanied by an accumulation of cells with slightly reduced PI staining relative to untreated G2 cells. Thus, the combination therapy mediates anti-leukemia activity by multiple mechanisms, including abrogation of cell cycle progression and induction of cell death. Addition of MERTK/ROCKi therapy to doxorubicin or etoposide additively decreased cell density relative to chemotherapy alone in OCI-AML5 and KG-1 cultures (p<0.0001). Treatment with doxorubicin and MERTK/ROCKi therapy also decreased OCI-AML5 cell density compared to MERTK/ROCKi therapy alone (p<0.01). These data indicate that combination therapies targeting MERTK and ROCK1/2, alone or in conjunction with chemotherapy, may be particularly effective for treatment of AML and support further studies in animal models. Citation Format: Dawn E. Barnes, Xiaodong Wang, Stephen V. Frye, H. Shelton Earp, Deborah DeRyckere, Douglas K. Barnes. MERTK and ROCK inhibitors mediate synergistic AML cell death and enhance therapeutic activity in combination with cytotoxic chemotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6388.

Superparamagnetic Iron Oxide Nanoparticles Carrying Chemotherapeutics Improve Drug Efficacy in Monolayer and Spheroid Cell Culture by Enabling Active Accumulation.

Cytotoxic and cytostatic chemotherapeutics act by attacking rapidly dividing tumor cells, predominantly affecting malignant tissue and to a certain degree preserving healthy cells. Nonetheless, severe side effects are caused as quickly proliferating healthy cells such as hematopoietic precursors and mucous membranes are impaired as well. This limits the administered dose and eventually allows tumor cells to escape treatment. In order to increase intratumoral drug concentration and simultaneously reduce systemic side effects, nanoparticles have come into focus as drug carriers. The functionalization of superparamagnetic iron oxide nanoparticles (SPIONs) with chemotherapeutics such as mitoxantrone (MTO) enables targeted drug transport by using magnetic forces. Here, we investigate SPIONs consisting of individual iron oxide cores of 10 nm in diameter and a total hydrodynamic diameter of 53 ± 0.8 nm as a transporting system for MTO. Comparing the killing efficacy in monolayer cell culture and multicellular tumor spheroids of HT-29 cells, we show that spheroids tolerate considerably higher doses of nanoparticle-loaded MTO. Therefore, dose predictions from conventional monolayer cell cultures are often misleading for in vivo applications. This was true for both soluble and nanoparticle-bound MTO. Using flow chambers mimicking in vivo blood flow, we furthermore demonstrate that SPIONs can magnetically accumulate MTO. We conclude that SPIONs can function as an effective delivery platform to increase local drug concentrations, thereby potentially overcoming chemotherapy resistance of cells.

The NSD2 p.E1099K Mutation Is Enriched at Relapse and Confers Drug Resistance in a Cell Context–Dependent Manner in Pediatric Acute Lymphoblastic Leukemia

The NSD2 p.E1099K (EK) mutation is observed in 10% of acute lymphoblastic leukemia (ALL) samples with enrichment at relapse indicating a role in clonal evolution and drug resistance. To discover mechanisms that mediate clonal expansion, we engineered B-precursor ALL (B-ALL) cell lines (Reh, 697) to overexpress wildtype (WT) and EK NSD2, but observed no differences in proliferation, clonal growth, or chemosensitivity. To address whether NSD2 EK acts collaboratively with other pathways, we used short hairpin RNAs to knockdown expression of NSD2 in B-ALL cell lines heterozygous for NSD2 EK (RS4;11, RCH-ACV, SEM). Knockdown resulted in decreased proliferation in all lines, decreased clonal growth in RCH-ACV, and increased sensitivity to cytotoxic chemotherapeutic agents, although the pattern of drug sensitivity varied among cell lines implying that the oncogenic properties of NSD2 mutations are likely cell context specific and rely on cooperative pathways. Knockdown of both Type II and REIIBP EK isoforms had a greater impact than knockdown of Type II alone, suggesting that both SET containing EK isoforms contribute to phenotypic changes driving relapse. Furthermore, in vivo models using both cell lines and patient samples revealed dramatically enhanced proliferation of NSD2 EK compared with WT and reduced sensitivity to 6-mercaptopurine in the relapse sample relative to diagnosis. Finally, EK-mediated changes in chromatin state and transcriptional output differed dramatically among cell lines further supporting a cell context-specific role of NSD2 EK. These results demonstrate a unique role of NSD2 EK in mediating clonal fitness through pleiotropic mechanisms dependent on the genetic and epigenetic landscape. IMPLICATIONS: NSD2 EK mutation leads to drug resistance and a clonal advantage in childhood B-ALL.

Internalized Functional DNA Aptamers as Alternative Cancer Therapies.

Despite major advances, cancer remains one of the largest burdens of disease worldwide. One reason behind this is that killing tumor cells without affecting healthy surrounding tissue remains a largely elusive prospect, despite the widespread availability of cytotoxic chemotherapeutic agents. To meet these modern healthcare requirements, it is essential to develop precision therapeutics that minimise off-target side-effects for various cancer types. To this end, highly specific molecular targeting agents against cancer are of great interest. These agents may work by targeting intracellular signalling pathways following receptor binding, or via internalization and targeting to specific subcellular compartments. DNA aptamers represent a promising molecular tool in this arena that can be used for both specific cell surface targeting and subsequent internalization and can also elicit a functional effect upon internalization. This review examines various cancer targeting cell-internalizing aptamers, with a particular focus towards functional aptamers that do not require additional conjugation to nanoparticles or small molecules to elicit a biological response. With a deeper understanding and precise exploitation of cancer specific molecular pathways, functional intracellular DNA aptamers may be a powerful step towards more widespread development of precision therapeutics.

Trispecific natural killer cell nanoengagers for targeted chemoimmunotherapy.

Activation of the innate immune system and natural killer (NK) cells has been a key effort in cancer immunotherapy research. Here, we report a nanoparticle-based trispecific NK cell engager (nano-TriNKE) platform that can target epidermal growth factor receptor (EGFR)-overexpressing tumors and promote the recruitment and activation of NK cells to eradicate these cancer cells. Moreover, the nanoengagers can deliver cytotoxic chemotherapeutics to further improve their therapeutic efficacy. We have demonstrated that effective NK cell activation can be achieved by the spatiotemporal coactivation of CD16 and 4-1BB stimulatory molecules on NK cells with nanoengagers, and the nanoengagers are more effective than free antibodies. We also show that biological targeting, either through radiotherapy or EGFR, is critical to the therapeutic effects of nanoengagers. Last, EGFR-targeted nanoengagers can augment both NK-activating agents and chemotherapy (epirubicin) as highly effective anticancer agents, providing robust chemoimmunotherapy.

Irinotecan-Associated Dysarthria in Patients with Pancreatic Cancer: A Single Site Experience

Case seriesPatients: Female, 56-year-old • Female, 72-year-old • Female, 68-year-oldFinal Diagnosis: Dysarthria asscoaited with irinotecan infusionSymptoms: Slurred speachMedication: —Clinical Procedure: —Specialty: OncologyObjective:Rare diseaseBackground:Irinotecan, a topoisomerase I inhibitor, is a cytotoxic chemotherapeutic agent used to treat multiple malignancies, including those of colorectal, pancreatic, cervical, esophageal, gastric, and lung origin. Dysarthria, a state of difficult or unclear articulation of speech, has been reported as a rare side effect of irinotecan through multiple case reports and case series, but with limited published data aimed at understanding the underlying mechanism and effective management strategies.Case Reports:We describe herein 3 cases of patients with pancreatic malignancy who experienced dysarthria while being treated with a chemotherapy regimen containing irinotecan at an ambulatory outpatient satellite chemotherapy site. All patients described received first-line FOLFIRINOX for pancreatic cancer and experienced dysarthria during their first infusion of irinotecan. In all cases, dysarthria was observed as a transient adverse drug reaction within the first 10 to 70 min of irinotecan infusion, which resolved rapidly upon pausing infusion without any long-term sequalae.All patients remained conscious and alert; physical and neurological examinations at dysarthria onset revealed no abnormalities. Some patients experienced distal extremity paresthesia, a known manifestation of oxaliplatin-induced acute neurotoxicity, and diaphoresis and nausea. Increased infusion time effectively prevented dysarthria during subsequent infusions.Conclusions:Oncologists, pharmacists, nurses, and other care team members should be aware that irinotecan-associated dysarthria is a rare, mild, and self-limiting phenomenon to avoid inadvertently altering or withholding therapy. We suggest extending irinotecan infusion time, as opposed to dose reduction or treatment discontinuation, as a practical clinical management strategy for patients who develop recurrent dysarthria secondary to irinotecan infusion.

The Potential of Tumor Debulking to Support Molecular Targeted Therapies.

Tumors may consist of billions of cells, which in malignant cases disseminate and form distant metastases. The large number of tumor cells formed by the high number of cell divisions during tumor progression creates a heterogeneous set of genetically diverse tumor cell clones. For cancer therapy this poses unique challenges, as distinct clones have to be targeted in different tissue locations. Recent research has led to the development of specific inhibitors of defined targets in cellular signaling cascades which promise more effective and more tumor-specific therapy approaches. Many of these molecular targeted therapy (MTT) compounds have already been translated into clinics or are currently being tested in clinical studies. However, the outgrowth of tumor cell clones resistant to such inhibitors is a drawback that affects specific inhibitors in a similar way as classical cytotoxic chemotherapeutics, because additionally acquired genetic alterations can enable tumor cells to circumvent the particular regulators of cellular signaling being targeted. Thus, it might be desirable to reduce genetic heterogeneity prior to molecular targeting, which could reduce the statistical chance of tumor relapse initiated by resistant clones. One way to achieve this is employing unspecific methods to remove as much tumor material as possible before MTT, e.g., by tumor debulking (TD). Currently, this is successfully applied in the clinical treatment of ovarian cancer. We believe that TD followed by treatment with a combination of molecular targeted drugs, optimally guided by biomarkers, might advance survival of patients suffering from various cancer types.

DBDx-based drug combinations show highly potent therapeutic efficacy against human pancreatic cancer xenografts in athymic mice

ABSTRACT Previous studies have shown that DBDx, a combination consisting of dipyridamole, bestatin and dexamethasone is highly effective against several cancer xenografts in athymic mice. Here the therapeutic effects of DBDx and its combination with gemcitabine or capcitabine against human pancreatic cancer xenografts and the mechanism were studied. In vivo experiments performed in athymic mice showed that the antitumor efficacy of DBDx was much stronger than that of gemcitabine or capecitabine alone. Notably, the combination of DBDx and gemcitabine or capcitabine further enhanced the efficacy. In the case of DBDx (242 mg/kg) plus gemcitabine (100 mg/kg), tumor weight decreased about 97.7%, and tumor sizes were shrinking during the treatment. In the case of DBDx (242 mg/kg) plus capecitabine (718.7 mg/kg), tumor weight decreased about 94.9%. Moreover, DBDx and its combinations obviously prolonged theoverall survival of mice compared with gemcitabine or capcitabine alone. DBDx-based drug combination therapy showed no obvious systematic toxicity. The gene expression profile analysis showed that the genes changed by DBDx were related to immune system and tumor vasculature. The result of protein array showed that the changed proteins in the serum of treated mice were related to immune and inflammation system. These results show that DBDx-based drug combinations, a new strategy which integrates the use of low-cytotoxic drugs and cytotoxic chemotherapeutics, are highly effective regimens against human pancreatic cancer in athymic mice at well tolerated doses. DBDx-based drug combination therapy might provide new options for the treatment of pancreatic cancer.

5-FU-Induced Upregulation of Exosomal PD-L1 Causes Immunosuppression in Advanced Gastric Cancer Patients.

Although the cytotoxic chemotherapeutic agent 5-fluorouracil (5-FU) is generally considered to directly kill cancer cells and exert immunostimulatory effects in advanced gastric cancer, accumulating evidence indicates that it upregulates the expression of PD-L1, a representative immune checkpoint blockade molecule involved in negative regulation of the immune response. It was reported that exosomes could transfer functional PD-L1 locally and distantly to suppress the antitumor immune response. However, whether 5-FU alters the expression of exosomal PD-L1 and induces immunosuppression in gastric cancer remains unclear. Herein, we found that 5-FU increased gastric cancer-derived exosomal PD-L1. Importantly, compared with baseline levels, circulating exosomal PD-L1 was significantly upregulated in 21 stage III–IV gastric cancer patients after two, four, and six repeated cycles of fluoropyrimidine treatment (P = 0.009, P = 0.047, and P = 0.023, respectively), accompanied by decreased amounts of IFN-γ, TNF-α, IL-2, IL-6, and GM-CSF (P = 0.014, P = 0.004, P = 0.009, P = 0.031, and P = 0.014, respectively). Additionally, circulating exosomal PD-L1 was increased more significantly in clinical non-responders compared with responders (P = 0.018). Furthermore, exosomal PD-L1 induced apoptosis in Jurkat T cells and inhibited T cell activation in PBMCs, which could be partly reversed by nivolumab. These results suggested that 5-FU-induced upregulation of exosomal PD-L1 causes systemic immunosuppression in advanced gastric cancer following multiple cycles of chemotherapy, especially after two cycles.

Triple-negative breast cancer: challenges and treatment options

TNBCs or Triple negative breast cancers are characterized by the deficiency of progesterone and estrogen receptors and also the absence down regulation of Human epithelial growth receptor type 2 (HER2). TNBCs have low prognosis rate because of heterogeneity.The heterogeneous nature of this cancer has constrained the effective progress in drug targeting among certain people.. In general HER2, PR and ER and the rate of proliferation are main predictive and prognostic factors in the detection of cancer of breast.Several pathways are involved in the progression of triple negative breast cancer from basal like cancer cells. The foremost being the loss by BRCA1-mediated pathway or mutation in the expression of several receptors.Certain groups have made some progress in unwinding TNBC's biological diversity and relating patterns of gene expression to molecular or genotypic subtypes.Earlier molecular categories of breast cancer use PAM50 via gene expression analysis to separate the breast cancer into the 4 intrinsic subtypes classified among many TNBCs in basal (BL) group and others divided between HER2 and luminal rich group. Currently, targeted therapy for TNBCs has not been approved. Nonetheless, a continuous progress has been made to detect the tumors at specific site for targeting and establish novel improved therapy.This review speaks about different approaches to TNBC treatment like cytotoxic therapy, targeted strategies, and chemotherapeutics by damage to DNA and targeting for repair of DNA and potent Nano carriers for targeting TNBC.

An experimental study of Ehrlich’s ascetic carcinoma in Swiss albino mice with chemotherapy

Background: Ehrlich-Lettre ascites carcinoma (EAC) is also known as Ehrlich cell. It was originally established as an ascites tumor in mice. The current concept that cytotoxic chemotherapeutic agents are administered at a dose to the maximum a patient can tolerate before the onset of severe and even life-threatening toxicity is still in wide clinical use. This study was conducted to evaluate the response of chemotherapy in the treatment of Ehrlich’s ascetic carcinoma. The various parameters to assess the response of various therapeutic schedules were regression of tumor by decrease in body weight of mice and decrease in abdominal girth; cell count of ascetic fluid and morphological changes of tumor cells after treatment with drugs and to study the percentage viability of the cells. Materials & Methods: A mouse bearing the tumor strain was taken from our laboratory in the Department of Pathology, IPGMER, Kolkata, where the strain was being maintained serially by inoculation of malignant cells into healthy mice every 8-10 days. In our work altogether 25 mice were taken for each set of experimental work. They were divided in four groups of 5-10 mice in each group. Results: All the mice in Group III revealed extensive metastasis with haemorrhagic ascetic fluid. About 60% mice of Group IV showed metastatic but there was no haemorrhagic fluid. None of the mice of Group IV survived beyond 35 days. Group III mice survived for a maximum of 25 days. Most of them died between 2-24 days. Conclusion: With the chemotherapy there was reduced cell count and cell viability. Actinomycin D alone produced a moderate effect on tumor cells.

Histological & Immunohistochemical Study on the Effect of Methotrexate on the Cerebellum of Adult Male Albino Rats and The Possible Protective Role of Lepidium Sativum

Introduction: Methotrexate is a cytotoxic chemotherapeutic agent, vastly utilized not only for malignancies but also for various inflammatory diseases, but it had hazardous effects on cerebellum. Lepidium sativum aherbal medication seemed to have neuroprotective effects.Objectives: This study was done to assess the effects of methotrexate on the cerebellum of adult male albino rats and the possible neuroprotective effect of lepidium sativum.Material and Methods: Sixty (60) adult male albino rats, weighing nearly 170-200gms were used in the current study. The first group (n=10) was the control group. Rats of second group(n=10)were given lepidium sativum aqueous extract (150mg/kg/day) by gastric tube daily for 4 weeks. Rats of third group (n=20)were given methotrexate (20mg/kg) on the 14th day then left without treatment for 2 weeks. Rats of fourth group (n=20)were given both Methotrexate and Lepidium Sativum as groups II and III. After 4 weeks, animals were sacrificed then the cerebellum was excised and processed for histological, immunohistochemical and quantitative studies.Results: Rats given lepidium sativum aquous extract was nearly similar to control group. Methotrexate treated rats showed significant decrease in number of Purkinje cells, significant increase in mean area % of GFAP positive immuno-staining and destructive changes of Purkinje cells with marked reduction of Nissl's granules. This was confirmed by electron microscopic examination and strong positive reaction for GFAP and caspase -3 reactions. Rats treated by Methotrexate and Lepidium Sativum showed nearly normal histological appearance of Purkinje cells with less vacuolated cytoplasm. This was confirmed by a significantrise in the number of Purkinje cells, significant diminution in caspase-3 positive cells and in GFAP immunostaining.Conclusion: Lepidium Sativum aqueous extract ameliorates the neurotoxic effects of methotrexate. So, it is recommended to use Lepidium Sativum, whenever methotrexate treatment is needed.