Presence Of Paclitaxel Research Articles

RBN-2397, a PARP7 Inhibitor, Synergizes with Paclitaxel to Inhibit Proliferation and Migration of Ovarian Cancer Cells.

Mono(ADP-ribosyl)ation (MARylation), a post translational modification of proteins, is emerging as an important regulator of the biology of cancer cells. PARP7 (TiPARP), a mono (ADP-ribosyl) transferase (MART), MARylates its substrate α-tubulin in ovarian cancer cells, promoting destabilization of microtubules, cell growth, and migration. Recent development of RBN-2397, a potent inhibitor that selectively acts on PARP7, has provided a new tool for exploring the role of PARP7 catalytic activity in biological processes. In this study, we investigated the role of PARP7 catalytic activity in the regulation of ovarian cancer cell biology via MARylation of α-tubulin. Ovarian cancer cell lines (OVCAR4, OVCAR3) were treated with RBN-2397 and paclitaxel, both separately and in combination. Western blotting and immunoprecipitation confirmed the effects of RBN-2397 on α-tubulin MARylation and stabilization. Cell proliferation and migration were assessed, and α-tubulin stabilization was quantified using immunofluorescent imaging. RNA-sequencing was performed to assess the effects on gene expression changes. RBN-2397 inhibited PARP7 activity, decreasing α-tubulin MARylation, leading to its stabilization, and reducing cancer cell proliferation and migration. The addition of paclitaxel further enhanced these effects, highlighting a synergistic interaction between the two drugs. Mutating the site of PARP7-mediated MARylation on α-tubulin similarly resulted in microtubule stabilization and decreased cell migration in the presence of paclitaxel. This study demonstrates that targeting PARP7 with RBN-2397, particularly in combination with paclitaxel, offers an effective strategy for inhibiting aggressive ovarian cancer cell phenotypes. Our findings underscore the potential of combining PARP7 inhibitors with established chemotherapeutics to enhance treatment efficacy in ovarian cancer.

Endophytic Aspergillus fumigatiaffinis: Novel paclitaxel production and optimization insights

This study investigated the potential of endophytic fungi to produce paclitaxel (Taxol®), a potent anticancer compound widely employed in chemotherapy. This research aimed to identify, confirm, and characterize endophytic fungi capable of paclitaxel (PTX) production and assess their paclitaxel yield. Additionally, it aimed to investigate factors influencing paclitaxel production. A total of 100 endophytic fungal isolates were collected and identified from the roots of Artemisia judaica. Aspergillus fumigatiaffinis exhibited the highest PTX production (26.373 μg L−1) among the isolated endophytic fungi. The strain was identified as A. fumigatiaffinis (Accession No. PP235788.1). Molecular identification confirmed its novelty, representing the first report of PTX production by A. fumigatiaffinis, an endophyte of Artemisia judaica. Optimization through full factorial design of experiments (DOE) and response surface methodology (RSM) significantly enhanced PTX production to 110.23 μg L−1 from 1 g of dry weight of the fungal culture under optimal conditions of pH 8.0, 150 μg L−1 becozyme supplementation, and 18 days of fermentation in potato dextrose broth. The presence of paclitaxel was confirmed using thin layer chromatography, high performance liquid chromatography, and gas chromatography–mass spectrometry. These findings maximize the role of endophytic fungus to produce a secondary metabolite that might be able to replace the chemically produced PTX and gives an opportunity to provide a sustainable source of PTX eco-friendly at high concentrations.Key points• Endophytic fungi, like A. fumigatiaffinis, show promise for eco-friendly paclitaxel production• Optimization strategies boost paclitaxel yield significantly, reaching 110.23 μg L−1• Molecular identification confirms novelty, offering a sustainable PTX source

Exosome-Mediated Transfer of ALDH2 in Nasopharyngeal Carcinoma Cells Confers Increased Resistance to Paclitaxel Treatment.

Nasopharyngeal carcinoma (NPC) is an aggressive and highly metastatic malignant tumor. Despite recent therapeutic advances, resistance to Taxol (the generic name of paclitaxel) therapy remains a major challenge in clinical management. Therefore, it is imperative to explore the potential mechanisms of paclitaxel resistance in NPC. This study aimed to investigate the expression of aldehyde dehydrogenase 2 (ALDH2) in NPC cells and its critical role in paclitaxel resistance. Paclitaxel-resistant cell line CNE1/Taxol (CNE1-TR), a drug-resistant cell line, was established by exposing the CNE1 nasopharyngeal carcinoma cell line to progressively increasing concentrations of paclitaxel. Furthermore, we investigated the role of ALDH2 in paclitaxel resistance and the function of exosomes using cell culture, Western blotting, reverse transcription-polymerase chain reaction (RT-PCR), Cell Counting Kit-8 (CCK-8), and nanoparticle tracking analysis. The results showed that in the presence of paclitaxel, the CNE1-TR cells manifested higher survival rate and half-maximal inhibitory concentration (IC50) value compared to the parental cell line, indicating strong resistance to paclitaxel. CNE1-TR cells had significantly upregulated mRNA and protein levels of ALDH2. In addition, exosome analysis showed that CNE1-TR cells were able to deliver ALDH2 via exosomes, increasing paclitaxel resistance in the recipient cells. We observed that the ALDH2 expression levels and paclitaxel resistance in CNE1-TR cells were effectively reduced by blocking the release of exosomes. ALDH2 is not only a key molecular marker indicative of therapeutic efficacy, but also a potential therapeutic target for developing novel anticancer strategies. By blocking the exosomal transport of ALDH2 or directly inhibiting its activity, it may be possible to overcome paclitaxel resistance, thus improving the success rate of clinical treatment.

Sonic hedgehog restrains the ubiquitin-dependent degradation of SP1 to inhibit neuronal/glial senescence associated phenotypes in chemotherapy-induced peripheral neuropathy via the TRIM25-CXCL13 axis

IntroductionChemotherapy-induced peripheral neuropathy (CIPN) is a common complication that affects an increasing number of cancer survivors. However, the current treatment options for CIPN are limited. Paclitaxel (PTX) is a widely used chemotherapeutic drug that induces senescence in cancer cells. While previous studies have demonstrated that Sonic hedgehog (Shh) can counteract cellular dysfunction during aging, its role in CIPN remains unknown. ObjectivesHerein, the aim of this study was to investigate whether Shh activation could inhibits neuronal/glial senescence and alleviates CIPN. MethodsWe treated ND7/23 neuronal cells and RSC96 Schwann cells with two selective Shh activators (purmorphamine [PUR] and smoothened agonist [SAG]) in the presence of PTX. Additionally, we utilized a CIPN mouse model induced by PTX injection. To assess cellular senescence, we performed a senescence-associated β-galactosidase (SA-β-gal) assay, measured reactive oxygen species (ROS) levels, and examined the expression of P16, P21, and γH2AX. To understand the underlying mechanisms, we conducted ubiquitin assays, LC-MS/MS, H&E staining, and assessed protein expression through Western blotting and immunofluorescence staining. ResultsIn vitro, we observed that Shh activation significantly alleviated the senescence-related decline in multiple functions included SA-β-gal activity, expression of P16 and P21, cell viability, and ROS accumulation in DRG sensory neurons and Schwann cells after PTX exposure. Furthermore, our in vivo experiments demonstrated that Shh activation significantly reduced axonal degeneration, demyelination, and improved nerve conduction. Mechanistically, we discovered that PTX reduced the protein level of SP1, which was ubiquitinated by the E3 ligase TRIM25 at the lysine 694 (K694), leading to increased CXCL13 expression, and we found that Shh activation inhibited PTX-induced neuronal/glial senescence and CIPN through the TRIM25-SP1-CXCL13 axis. ConclusionThese findings provide evidence for the role of PTX-induced senescence in DRG sensory neurons and Schwann cells, suggesting that Shh could be a potential therapeutic target for CIPN.

Synchrotron microtomography reveals insights into the degradation kinetics of bio-degradable coronary magnesium scaffolds

Bioresorbable magnesium scaffolds are a promising future treatment option for coronary artery stenosis, especially for young adults. Due to the degradation of these scaffolds (<1 year), long-term device-related clinical events could be reduced compared to treatments with conventional drug eluting stents. First clinical trials indicate a return of vasomotion after one year, which may be associated with improved long-term clinical outcomes. However, even after decades of development, the degradation process, ideal degradation time and biological response in vivo are still not fully understood.The present study investigates the in vivo degradation of magnesium scaffolds in the coronary arteries of pigs influenced by different strut thicknesses and the presence of antiproliferative drugs. Due to high 3D image contrast of synchrotron-based micro-CT with phase contrast (SR-μCT), a qualitative and quantitative evaluation of the degradation morphology of magnesium scaffolds was obtained. For the segmentation of the μCT images a convolutional network architecture (U-net) was exploited, demonstrating the huge potential of merging high resolution SR-μCT with deep learning (DL) supported data analysis.In total, 30 scaffolds, made of the rare earth alloy Resoloy®, with different strut designs were implanted into the coronary arteries of 10 domestic pigs for 28 days using drug-coated or uncoated angioplasty balloons for post-dilatation. The degradation morphology was analyzed using scanning electron microscopy, energy dispersive x-ray spectroscopy and SR-μCT. The data from these methods were then related to data from angiography, optical coherence tomography and histology.A thinner strut size (95 vs. 130 μm) and the presence of paclitaxel indicated a slower degradation rate at 28 d in vivo, which positively influences the late lumen loss (0.5 and 0.6 mm vs. 1.0 and 1.1 mm) and recoil values (0 and 1.7% vs. 6.1 and 22%).

Comparative Evaluation of Two Paclitaxel-Coated Stents in an Experimental Setting.

Unlike paclitaxel-coated balloons, pre-clinical data comparing different paclitaxel-coated stents (PCSs) are weak. The study objective was to compare the features of the 2 main PCSs: Eluvia® (Boston Scientific, Marlborough, MA) versus ZilverPTX® (Cook Medical, Bloomington, IN). Analysis was carried out on 12 pigs divided into 2 groups: Eluvia® (n=6) and ZilverPTX® (n=6). The pigs received the PCS corresponding to their group in each external iliac artery and were paired one by one, to examine 6 different post-implantation timepoints: after 30 minutes, 6 hours, 24 hours, 3 days, 7 days, and 14 days. The paclitaxel concentration measurements and the histological analysis were carried out under blind testing on the plasma, arterial, lymph node, and muscle samples. A linear regression model and Wilcoxon Mann-Whitney test were used to study the variables. The plasma paclitaxel rate decrease over 24 hours after PCS implantation was significantly different between the two groups, expressed by the correlation coefficient 0.19 (0.14-0.23; p<0.001) with an undetectable concentration at the 10th hour for Eluvia® versus 3 days for ZilverPTX®. Significantly higher paclitaxel concentrations with ZilverPTX® PCS were observed in muscle samples at each timepoint: extensor digitorum brevis 3.2 (1.17-5.23; p=0.005), biceps femoris 4.27 (2.27-6.26; p<0.001), semi-tendinosus 3.79 (1.85-5.73; p=0.001), tibialis anterior 3.0 (1.37-4.64; p=0.001), and in the femoral nodes 2.27±1.74 ng/g versus 0.14±0.13 ng/g (p<0.001). Histological analysis revealed a trend for more marked intimal inflammation in the arteries stented with ZilverPTX® (p=0.063), especially after the 7th and 14th days. Such a difference in the concentration of paclitaxel in the plasma, muscles, and lymph nodes between the two stents was higher than expected based on differences in device design. The clinical consequences of these results remain to be elucidated, particularly regarding the concerning presence of paclitaxel in muscles and adjacent lymph nodes. This experimental study compares 2 paclitaxel-coated stents. It demonstrates that differences in stent designs and drug features (coatings and concentrations) translate into differences in terms of concentrations of paclitaxel in the plasma, muscles, and lymph nodes. Our results favor the Eluvia® stent over the ZilverPTX® stent, although more studies are required to confirm this conclusion.

The first facile optical density-dependent approach for the analysis of doxorubicin, an oncogenic agent accompanied with the co-prescribed drug; paclitaxel

Doxorubicin (DRB) is an anthracycline oncogenic drug extracted from cultures of Streptomyces peucetius var. caesius. It is frequently recommended as an anti-neoplastic agent for the treatment of diverse malignancies. It exerts its antineoplastic effect either via inhibiting the enzyme topoisomerase II and/or via intercalation to DNA or reactive oxygen species generation. In the present article, the direct, simple, one-pot, somewhat eco-safe, and non-extractive spectrophotometric system was executed to track doxorubicin, a chemotherapeutic remedy, in the presence of paclitaxel, a naturally occurring Taxan antineoplastic radical, through the greenness rated method. DRB’s optical density was studied in various mediums and solvents to develop the current approach. An acidic ethanolic solution was found to increase the optical density of the sample significantly. At 480 nm., the most remarkable optical density was obtained. Various experimental factors, including intrinsic media, solvent, pH, and stability time, were investigated and controlled. The current approach achieved linearity within the 0.6–40.0 µg mL-1 range, accompanied by a limit of both detection and quantification (LOD and LOQ) of 0.18 and 0.55 µg mL-1, correspondingly. The approach was validated under the ICH guidelines (Quality Guidelines). The system’s greenness and enhancement degree were estimated.

Abstract 1284: The effect of Semaphorin 7A on breast cancer metastases and chemoresistance in young women

Abstract Our overall goal is to understand drivers of chemoresistance in triple-negative breast cancer (TNBC) and the mechanisms that contribute to metastatic spread in young women. TNBC is more frequently diagnosed in women aged &amp;lt;40 and confers worse overall and distant-metastatic-free survival. Our lab identified a ß1 integrin-ligand, Semaphorin 7A (SEMA7A), that is increased in TNBC and young women's breast cancer (BC). Our central hypothesis is that SEMA7A is increased in TNBC and signals via ß1-integrin to activate epithelial-to-mesenchymal plasticity (EMP) and chemoresistance. We have incorporated genetically manipulated TNBC cell lines and ex vivo tumor samples to define how SEMA7A expression impacts metastasis and chemoresistance. We have also incorporated our Young Women’s BC Cohort to analyze SEMA7A protein expression in human tumors and mammary epithelial tissue. In this cohort, we found that the normal mammary epithelium and tumors exhibit higher SEMA7A expression in women with postpartum BC, defined as BC diagnosed within 10 years postpartum, who are between 30&amp;39-years-old. This sub-population also has increased tumor-associated lymphatic vessel density, as well as increased lymphovascular invasion and LN involvement. Using a 9-month-old mouse model, which is equivalent to a 35-year-old woman, we have shown that mammary tumor growth and metastasis are increased compared to young 8-week-old mice, which can be mimicked by SEMA7A OE tumors in the young mice; these results suggest that the aged microenvironment can replicate the tumor promotional effects of SEMA7A. Similarly, in orthotopic models, SEMA7A OE cells metastasize more frequently to lymph nodes (LNs) and lungs compared to controls. Additionally, via a tail vein injection, we observe that SEMA7A OE cells survive in circulation, seed the lung, and outgrow faster compared to controls. In vitro, SEMA7A promotes migration and invasion via ß1-integrin signaling. Finally, ex vivo metastatic tumor cells, when compared to parental cells, exhibit evidence of EMP. Namely, prior to dissemination, SEMA7A OE cells are mostly mesenchymal-like. Ex vivo, SEMA7A OE cells from LN metastasis express both epithelial/mesenchymal markers, suggesting a hybrid phenotype and maintenance of an invasive phenotype. Whereas those from the lung are more epithelial, which can be more proliferative. EMP is also known to promote chemoresistance and we have found that SEMA7A OE cells are resistant to caspase-mediated cell death in the presence of paclitaxel, a known chemotherapeutic utilized for TNBC, and have increased activation of pAkt, which can mediate pro-survival phenotypes. Overall, our results suggest that the aged mammary gland and SEMA7A expression can promote metastatic spread and chemoresistance and that these patient populations should be considered for direct and indirect targeting of SEMA7A by novel therapeutics. Citation Format: Kelsey Kines, Sarah Tarullo, Virginia Borges, Traci Lyons. The effect of Semaphorin 7A on breast cancer metastases and chemoresistance in young women [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1284.

Screening and characterization of paclitaxel-producing fungus \(\textit{Talaromyces wortmannii}\) WQF18 isolated from \(\textit{Cephalotaxus mannii }\)Hook. f..

Cephalotaxus mannii Hook. f. is a rare evergreen conifer native listed in the International Union for Conservation of Nature (IUCN) Red List, which is utilized for leukemia treatment. Although endophytic fungi from C. mannii was reported before, their cytotoxic property has not been revealed yet. In the present study, a total of 7 endophytic fungi were isolated from C. mannii collected in Ha Giang province, Vietnam, among which the isolate WQF18 was active against 5 tested pathogens with inhibition zones ranging from 18.0 ± 0.7 to 25.0 ± 0.4 mm. In addition, only ethyl acetate extract of isolate WQF18 showed cytotoxicity on A549 and MCF7 cell lines with IC50 values of 69.6 ± 2.3 µg/mL and 78.6 ± 1.6 µg/mL, respectively. PCR-based molecular screening revealed that the positive hits for both 10-deacetylbaccatin III-10-O-acetyl transferase (dbat) and taxadiene synthase (ts) genes involved in paclitaxel biosynthesis were only observed in the WQF18 isolate. Based on morphological and molecular identification, the WQF18 isolate was identified as Talaromyces wortmannii WQF18. The presence of paclitaxel in T. wortmannii WQF18 was further confirmed by dbat sequence alignment, phenotypic, and HPLC-DAD analysis. To the best of our knowledge, this is the first report demonstrating the paclitaxel-producing capability of endophytic fungi from C. mannii. These findings provide a new platform for deciphering paclitaxel biosynthesis of endophytic fungi from non-Taxus plants and further paclitaxel production.

Abstract 2734: Integrating hybrid spatiotemporal models and multiscale data for the study of cancer progression in 3D cultures

Abstract Tumour progression consists of various stages including migration to the surrounding tissues, leading to metastasis [1]. In this work, we investigate the multiscale quantitative characteristics of the spatial organization and migration of cancer in 3D cultures. For this study, we combined 3D cell culture experiments of Triple Negative Breast Cancer (TNBC) cells, hybrid spatiotemporal models, and longitudinal RNA-sequencing data analysis. The experiments included two experimental conditions; simple growth experiments, and experiments of growth in presence of the migrastatic drug Paclitaxel. The spatial distributions of the cells enabled us to calibrate and validate a hybrid Keller-Segel model, incorporated into a novel computational framework capable of interpreting the relation between morphological patterns and the underlying mechanisms of cancer growth [2]. The RNA-seq data included different time-points with and without treatment. The results suggested that cancer cells exhibited biased movement towards the bottom of the space, a movement that was inhibited in the presence of Paclitaxel. The calibrated model was able to describe the overall characteristics of the experimental observations, and suggested that cancer cells exhibited chemotactic migration and cell accumulation, as well as random motion throughout the period of development. The spatial pattern analysis revealed transient, non-random spatial distributions of cancer cells that consisted of clustered patterns across a wide range of neighbourhood distances, as well as dispersion for larger distances. The RNA-seq data exhibited coherence with the chemotactic migration hypothesis of the mathematical model, indicating significant under-representation of the Gene Ontology (GO) terms related to chemotactic migration in presence of the migrastatic drug. Overall, this study provided an insightful quantitative characterization of the spatiotemporal organization and progression of TNBC cells in 3D cultures. We anticipate that these developments will enable us to expand our studies to more realistic conditions, including the introduction of heterogeneic cell populations.

Abstract 3127: Negative impact of paclitaxel on human breast tumor microenvironment and its reversal by the combination of interferon-α with TLR3 agonist rintatolimod

Abstract Background: Taxanes are extensively used in the chemotherapy of breast cancer in the neoadjuvant, adjuvant and metastatic settings. Paclitaxel is known to have both immunostimulatory and immunosuppressive effects, but its impact on the chemokine system within the tumor microenvironment (TME) remains unclear. In the breast TME treated with neoadjuvant taxane chemotherapy, high levels of the cytotoxic T cell (CTL) attractants CXCL9, CXCL10 and their common receptor CXCR3 predict higher probability of pathological complete response (pCR) and improved long term outcomes. We previously developed a chemokine modulatory regimen (CKM) combining interferon-α with TLR3 agonist rintatolimod to selectively increase CTL-attracting chemokines in the TME but reduce the production of chemokines attracting myeloid-derived suppressor cells (MDSC) and regulatory T cells (Treg) in the TME. Here we evaluated paclitaxel’s impact on chemokine production in the human breast TME and CKM’s ability to eliminate undesirable aspects of taxane chemotherapy. Methods: Fresh breast cancer tissues obtained during routine surgeries, breast cancer cell lines BT-549 and MDA-MD-231 and peripheral blood monocyte-derived macrophages were analyzed immediately or cultured ex vivo for 24 hours in the absence or presence of paclitaxel and/or CKM components. The expression of chemokine genes and secretion of chemokines by freshly harvested and ex vivo-treated tumor explants, cancer cell lines and macrophages were analyzed by quantitative PCR (Taqman) and ELISA. Paired student t-test was used for statistical analyses. Results: Breast cancer explants spontaneously expressed high levels of MDSC/Treg-attractants CXCL12 and CCL22, but only marginal levels of CTL and natural killer (NK) cell attractants CXCL9, CXCL10, CXCL11 and CCL5. Unexpectedly, paclitaxel treatment resulted in further elevation of granulocyte/MDSC-attractant CXCL8 and CCL22, which was reversed by the combination of paclitaxel with CKM. At the same time, while paclitaxel alone did not induce any of the CTL attractants tested, its combination with the CKM was highly effective in inducing CXCL9, CXCL10, CXCL11 and CCL5. Conclusions: Our results identify an undesirable aspect of paclitaxel’s impact on breast cancer. The ability of CKM to enhance the expression of CTL/NK cell attractants and suppress the production of Treg/MDSC attractants produced by paclitaxel provides a strong rationale for combined use of CKM in taxane-based chemo-immunotherapy of breast cancer and potentially other diseases. Citation Format: Shipra Gandhi, Melissa Grimm, Ronald Slomba, Kazuaki Takabe, Pawel Kalinski. Negative impact of paclitaxel on human breast tumor microenvironment and its reversal by the combination of interferon-α with TLR3 agonist rintatolimod [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3127.

Docosahexaenoic acid (DHA) and linoleic acid (LA) modulate the expression of breast cancer involved miRNAs in MDA-MB-231 cell line.

Docosahexaenoic acid (DHA) and linoleic acid (LA) have modulatory effects on breast cancer (BC) cell lines. We aimed to investigate the effects of DHA, LA alone, in combination, and in the presence of paclitaxel on the expression of five microRNAs involved in the pathology of BC in MDA-MB-231 cell line. MDA-MB-231cells were treated with either DHA or LA or in combination in the presence/absence of paclitaxel (Taxol). Total RNA was extracted and cDNA synthesized from the cells before and after treatment. The expression levels of miR-30, miR-106b, miR-20, miR-126, and miR-194 were determined by quantitative real-time PCR (qPCR). Treatment of MDA-MB-231cells with DHA modulated the gene expression of miR-30 (increased by 7.74-fold (p<0.0001), miR-194 (decreased by 11-fold (p<0.0001)), miR-106b (increased by 2.64-fold (p=0.0004), miR-126 (decreased by 50-fold (p<0.0001)), and miR-20 (decreased by 4-fold (p<0.0001)). Additionally, treatment of MDA-MB-231cells with LA modulated the gene expression of miR-30 (increased by 2.38-fold (p=0.0001)), miR-194 (decreased by 100-fold (p<0.0001)), miR-106b (decreased by 10-fold (p<0.0001)). The combined DHA/LA treatment of MDA-MB-231cells showed regulatory effect on the expression of studied microRNAs in which decreased the expression of miR-30 (5.5-fold (p<0.0001)), miR-194 (11-fold (p<0.0001)), miR-20 (3.5-fold (p=0.0006)), and increased the expression of miR-106b (9.78-fold (p<0.0001)). Modulation of the expression levels of BC-involved microRNAs could be one of the possible mechanisms of action through which DHA and LA may exert their biologic effects on MDA-MB-231cell line.

The effect of Chaihu-shugan-san on cytotoxicity induction and PDGF gene expression in cervical cancer cell line HeLa in the presence of paclitaxel +cisplatin.

Chaihu-shugan-san, as a traditional Chinese herbal formula, is composed of seven different herbs. This medicine can treat cancer due to its antioxidant compounds. In this study, the effect of Chaihu-shugan-san was considered on cytotoxicity induction and PDGF gene expression in cervical cancer cell line HeLa at different concentrations and at different times, by the MTT method. Paclitaxel + cisplatin were used as a control in this study. The expression of the PDGF gene was quantitatively evaluated in treated cells by real-time PCR, and a generalized linear model was used to evaluate the effect of the medicine, and Duncan's multiple range tests were used to evaluate the data. The results of the MTT test showed that Chaihu-shugan-san had antitumor properties in different concentrations, but there was a significant difference between this medicine and paclitaxel +cisplatin. Also, examination of gene expression showed that this medicine reduced the expression of the PDGF gene in the HeLa cancer cell line (P ? 0.04). Therefore, Chaihu-shugan-san could be suggested as an effective factor in preventing the growth of cervical cancer cells and controlling angiogenic factors that play an important role in the metastasis of cancerous tumors.

Single-Shot Local Injection of Microfragmented Fat Tissue Loaded with Paclitaxel Induces Potent Growth Inhibition of Hepatocellular Carcinoma in Nude Mice.

Simple SummaryTherapeutic approaches to increase localization of chemotherapy at the tumor site, reducing systemic toxicity, are under deep investigation. In previous studies, we have shown that microfragmented adipose tissue (MFAT) may act as a natural scaffold able to deliver anti-cancer drugs. We demonstrated that MFAT and its devitalized counterpart (DMFAT) are able to absorb significant amounts of the chemotherapeutic drug Paclitaxel (PTX), with the ability to kill many different human cancer cell lines in vitro and in vivo, preventing tumor relapse when placed in the surgical area of tumor resection. We demonstrated here for the first time that DMFAT loaded with PTX was also very effective in inhibiting the in vivo growth of hepatocellular carcinoma (HCC) in an advanced stage of progression, suggesting it as a new potent and viable drug-delivery system that may be hypothetically translated to treat inoperable primary tumors in humans.Hepatocellular carcinoma (HCC) is poorly beneficiated by intravenous chemotherapy due to inadequate availability of drugs at the tumor site. We previously demonstrated that human micro-fragmented adipose tissue (MFAT) and its devitalized counterpart (DMFAT) could be effective natural scaffolds to deliver Paclitaxel (PTX) to tumors in both in vitro and in vivo tests, affecting cancer growth relapse. Here we tested the efficacy of DMFAT-PTX in a well-established HCC in nude mice. MFAT-PTX and DMFAT-PTX preparations were tested for anti-cancer activity in 2D and 3D assays using Hep-3B tumor cells. The efficacy of DMFAT-PTX was evaluated after a single-shot subcutaneous injection near a Hep-3B growing tumor by assessing tumor volumes, apoptosis rate, and drug pharmacokinetics in an in vivo model. Potent antiproliferative activity was seen in both in vitro 2D and 3D tests. Mice treated with DMFAT-PTX (10 mg/kg) produced potent Hep-3B growth inhibition with 33% complete tumor regressions. All treated animals experienced tumor ulceration at the site of DMFAT-PTX injection, which healed spontaneously. Lowering the drug concentration (5 mg/kg) prevented the formation of ulcers, maintaining statistically significant efficacy. Histology revealed a higher number of apoptotic cancer cells intratumorally, suggesting prolonged presence of PTX that was confirmed by the pharmacokinetic analysis. DMFAT may be a potent and valid new tool for local chemotherapy of HCC in an advanced stage of progression, also suggesting potential effectiveness in other human primary inoperable cancers.

Effects of paclitaxel in mitochondrial function and cellular phenotype in human peripheral blood mononuclear cells and monocytes

Chemotherapy-induced neuropathy (CIPN) is a common complication of paclitaxel. CIPN affects the quality of life of cancer survivors and frequently leads to discontinuation of treatment. Paclitaxel affects neuronal microtubules and induces neuronal mitochondrial dysfunction. However, there is limited clinical information regarding paclitaxel's effects on monocytes. Preclinical studies suggest that paclitaxel-induced neuronal damage is driven by monocytes/macrophages. Therefore, we evaluated whether paclitaxel selectively induces mitochondrial dysfunction and a pro-inflammatory phenotype in human circulating monocytes. We conducted studies in human primary peripheral blood mononuclear cells (PBMCs) from cancer patients being treated with paclitaxel, and in vitro analysis in PBMC cells and monocytes, and THP-1 monocytes in the presence of paclitaxel (0.1, 1, 10 uM). We used flow cytometric markers to study mitochondrial reactive oxygen species (ROS) and mitochondrial membrane potential, namely MitoSox and DIOC6(3) respectively. We also measured mRNA levels of pro- and anti-inflammatory molecules using qRT-PCR. In vitro paclitaxel induced a depolarization state in mitochondria in THP-1, human primary monocytes, and primary human PBMCs, but it did not change MitoSox. Monocytes in PBMCs cells from patients treated with paclitaxel showed significative depolarization state in mitochondria when compared to cells from control patients. In THP-1 cells, paclitaxel enhanced mRNA levels of the pro-inflammatory cytokines IL-8 and TNF alpha. In human primary PBMCs, paclitaxel reduced the anti-inflammatory factors CD163 and IL-10, and enhanced the TNF alpha, COX-2 and MCP-1 mRNA levels. Our study provides evidence that paclitaxel can induce mitochondrial dysfunction in isolated human monocytes and in monocytes present in total PBMCs cells. The observed depolarizing changes are indicative of a pro-mitophagy state, which is in accordance with the paclitaxel-induced pro-inflammatory phenotype in these cells. Early detection of mitochondria dysfunction in human monocytes could be a predictable sign to CIPN development in cancer patients. Our research was supported by the Early-Career Investigator Award W81XWH-16-1-0438 of the Department of Defense, The Pershing Square Sohn Cancer Research Alliance, Weill Cornell Medicine Funds, Department of Anesthesiology-Wake Forest School of Medicine Funds, Comprehensive Cancer Center-Wake Forest School of Medicine Funds, NIDA R21CA248106, National Center for Advancing Translational Sciences (NCATS)-NIH through Grant Award Number UL1TR001420. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Biocompatible Ionic Liquid-Mediated Micelles for Enhanced Transdermal Delivery of Paclitaxel.

Chemotherapeutic cytotoxic agents such as paclitaxel (PTX) are considered essential for the treatment of various cancers. However, PTX injection is associated with severe systemic side effects and high rates of patient noncompliance. Micelle formulations (MFs) are nano-drug delivery systems that offer a solution to these problems. Herein, we report an advantageous carrier for the transdermal delivery of PTX comprising a new MF that consists of two biocompatible surfactants: cholinium oleate ([Cho][Ole]), which is a surface-active ionic liquid (SAIL), and sorbitan monolaurate (Span-20). A solubility assessment confirmed that PTX was readily solubilized in the SAIL-based micelles via multipoint hydrogen bonding and cation-π and π-π interactions between PTX and SAIL[Cho][Ole]. Dynamic light scattering (DLS) and transmission electron microscopy revealed that in the presence of PTX, the MF formed spherical PTX-loaded micelles that were well-distributed in the range 8.7-25.3 nm. According to DLS, the sizes and size distributions of the micelle droplets did not change significantly over the entire storage period, attesting to their physical stability. In vitro transdermal assessments using a Franz diffusion cell revealed that the MF absorbed PTX 4 times more effectively than a Tween 80-based formulation and 6 times more effectively than an ethanol-based formulation. In vitro and in vivo skin irritation tests revealed that the new carrier had a negligible toxicity profile compared with a conventional ionic liquid-based carrier. Based on these findings, we believe that the SAIL[Cho][Ole]-based MF has potential as a biocompatible nanocarrier for the effective transdermal delivery of poorly soluble chemotherapeutics such as PTX.

Targeted delivery of paclitaxel by NL2 peptide-functionalized on core-shell LaVO4: Eu3@ poly (levodopa) luminescent nanoparticles.

Targeted drug delivery enhances drug efficiency and selectivity without affecting normal cells. Luminescent nanoparticles can be used for tumor imaging as well as selective tumor targeting for drug delivery. In this research, LaVO4 :Eu3+ was synthesized, the luminescent nanocrystal was coated by surface polymerization of levodopa in the presence of Paclitaxel (PTX), and then NL2 peptide was coupled on the surface of polymer-coated luminescent nanoparticles. Next, the capability of the modified drug was examined by in vitro and in vivo experiments. MTT assay on SK-BR-3 cell line (as breast cancer cells) and fluorescent microscopy results indicate that this modification decreases significantly drug toxicity and increases its selectivity. In addition, in vivo experiments confirm more capability of the NL2-functionalized nanocomposite for reducing tumor size, drug distribution in the body, and more aggregation of PTX in tumor tissue. Overall, it is concluded that tumor imaging is possible using luminescent LaVO4 :Eu3+ core and NL2 peptide increases significantly the specificity of PTX in combination with a functionalized luminescent polymeric carrier.

Molecular subtyping and functional validation of TTK, TPX2, UBE2C, and LRP8 in sensitivity of TNBC to paclitaxel

Triple-negative breast cancer (TNBC) patients exhibit variable responses to chemotherapy, suggesting an underlying molecular heterogeneity. In the current study, we analyzed publicly available transcriptome data from 360 TNBC and 88 normal breast tissues, which revealed activation of nucleosome and cell cycle as the hallmarks of TNBC. Mechanistic network analysis identified activation of FOXM1 and ERBB2, and suppression of TP53 and NURP1 networks in TNBC. Employing Iterative Clustering and Guide-gene Selection (ICGS), Uniform Manifold Approximation and Projection (UMAP), and dimensionality reduction analyses, we classified TNBC into seven molecular subtypes, each exhibiting a unique molecular signature, including immune infiltration (CD19, CD8, and macrophages) and mesenchymal signature, which correlated with variable disease outcomes in a larger cohort (1,070) of BC. Mechanistically, depletion of TTK, TPX2, UBE2C, CDCA7, MELK, NFE2L3, DDX39A, and LRP8 led to substantial inhibition of colony formation of TNBC models, which was further enhanced in the presence of paclitaxel. Our data provide novel insights into the molecular heterogeneity of TNBC and identified TTK, TPX2, UBE2C, and LRP8 as main drivers of TNBC tumorigenesis.

Facile one-pot green solvent synergized fluorescence reaction for determination of doxorubicin in presence of paclitaxel; co-administered drug, application to stability study and analysis in bulk, vial and biological fluids

A green, fast, simple, economical, ultra-sensitive and accurate spectrofluorimetric method was described in the article for the estimation of Doxorubicin hydrochloride (DOX), a type II topoisomerase enzyme inhibitor either alone and in the presence of co- administered drug; paclitaxel as adjuvant chemotherapy combination in bulk, pharmaceutical dosage form and biological fluids. The proposed method was based on studying the fluorescence demeanor of DOX in different organized media and solvents. The intrinsic fluorescence was found to be greatly enhanced in acidic ethanolic solution. The maximum intensity of fluorescence was observed at 590 nm after excitation at 475 nm. Different experimental parameters were examined and adjusted involving various organized media, solvent, and pH. The methods was validated according to ICH guidelines. The linearity of the method was accomplished in the range of 7 × 10−4–4.0 µg ml−1. The values of LOD and LOQ were 0.2 and 0.6 ng ml−1 respectively. The present method is extremely sensitive enough for determination of traces of the drug in various biological fluids with excellent % recovery. The proposed method was extended to investigate the stability of DOX following various induced degradation protocols including: acidic, alkaline, oxidative and photolytic. In addition, the kinetics of the degradation of DOX was investigated and the apparent first-order rate constant, half-life times and Quantum yield were calculated.

Modulating TRPV4 channels with paclitaxel and lithium

Transient receptor potential V4 (TRPV4), a plasma membrane calcium channel, is implicated as a contributor to the initiation of chemotherapy-induced peripheral neuropathy (CIPN). Paclitaxel (PTX) is a commonly used anticancer drug that causes CIPN and lithium has been shown to prevent CIPN. However, the direct effect of PTX and lithium on TRPV4 is not clear. This study investigated these actions using biochemical, pharmacological, and electrophysiological approaches using a neuronal cell line (SH-SY5Y). The addition of pharmacologically appropriate levels of PTX increased the expression of TRPV4, TRPV4 currents, and TRPV4-dependent calcium fluxes. Prolonged exposure to PTX amplified the acute effects of TRPV4 expression, currents, and calcium fluxes. Pretreatment with lithium (1 mM) decreased TRPV4 currents and calcium fluxes in the absence and presence of PTX. These findings enhance our understanding of the properties and regulation of TRPV4, the cellular mechanisms of PTX-induced neuropathy, and the mechanism of lithium for prevention of CIPN.