Drug Repositioning Screen on a New Primary Cell Line Identifies Potent Therapeutics for Glioblastoma

Glioblastoma is a malignant brain cancer with limited treatment options and high mortality rate. While established glioblastoma cell line models provide valuable information, they ultimately lose most primary characteristics of tumors under long-term serum culture conditions. Therefore, established cell lines do not necessarily recapitulate genetic and morphological characteristics of real tumors. In this study, in line with the growing interest in using primary cell line models derived from patient tissue, we generated a primary glioblastoma cell line, KUGBM8 and characterized its genetic alterations, long term growth ability, tumor formation capacity and its response to Temozolomide, the front-line chemotherapy utilized clinically. In addition, we performed a drug repurposing screen on the KUGBM8 cell line to identify FDA-approved agents that can be incorporated into glioblastoma treatment regimen and identified Topotecan as a lead drug among 1,200 drugs. We showed Topotecan can induce cell death in KUGBM8 and other primary cell lines and cooperate with Temozolomide in low dosage combinations. Together, our study provides a new primary cell line model that can be suitable for both in vitro and in vivo studies and suggests that Topotecan can offer promise as a therapeutic approach for glioblastoma.

Primary tumor cell cultures: сurrent methods of obtaining and subcultivation

Over the past decades, transplantable cell lines have been an affordable model for studying the biology and effect of chemotherapeutic drugs on tumors. However, numerous studies have shown that these cell lines are not heterogeneous enough and cannot reflect the drug resistance of tumors that occurs in some patients. Primary cell line cultures isolated from solid tumors have become widespread in personalized cancer therapy. This review discusses the basic methods for the preparation and cultivation of primary cell lines. A brief description is given of the methods for the disaggregation of tumor material using enzymatic, chemical and mechanical dissociation. The systems of cultivation of primary cell cultures. The selection of an appropriate dissociation method and cultivation is important to preserve the benefits of primary culture in preclinical studies.

Usutu Virus Infection of Embryonated Chicken Eggs and a Chicken Embryo-Derived Primary Cell Line

Usutu virus (USUV) is a mosquito-borne flavivirus, closely related to the West Nile virus (WNV). Similar to WNV, USUV may cause infections in humans, with occasional, but sometimes severe, neurological complications. Further, USUV can be highly pathogenic in wild and captive birds and its circulation in Europe has given rise to substantial avian death. Adequate study models of this virus are still lacking but are critically needed to understand its pathogenesis and virulence spectrum. The chicken embryo is a low-cost, easy-to-manipulate and ethically acceptable model that closely reflects mammalian fetal development and allows immune response investigations, drug screening, and high-throughput virus production for vaccine development. While former studies suggested that this model was refractory to USUV infection, we unexpectedly found that high doses of four phylogenetically distinct USUV strains caused embryonic lethality. By employing immunohistochemistry and quantitative reverse transcriptase-polymerase chain reaction, we demonstrated that USUV was widely distributed in embryonic tissues, including the brain, retina, and feather follicles. We then successfully developed a primary cell line from the chorioallantoic membrane that was permissive to the virus without the need for viral adaptation. We believe the future use of these models would foster a significant understanding of USUV-induced neuropathogenesis and immune response and allow the future development of drugs and vaccines against USUV.

PSVIII-17 In vitro and in vivo investigations of effects of natural phytogenic compounds on muscle cell gene expressions and growth performance and carcass composition of finishing pigs

For lean meat production in swine industry, some synthetic compounds have been applied as feed additives in finishing swine feeds. However, this practice raises public concern due to potential imbalance of innate hormones coupled with a possible residue of the synthetic compounds in meat products after ingestion. In this study, we investigated natural phytogenic compounds (NPC) from fruit peel consisted of mainly ursolic and maslinic acids to enhance muscle development but decrease fat deposition in swine. To test effects on cell line, NPC was treated in the porcine primary cell line obtained from muscle tissue of piglets at approximately 6.5 kg of body weight and NIH-3T3-L1 cell line from mouse adipose tissue cell (Experiment 1). In the porcine primary cell line, immunofluorescence measurement indicated that myogenin expression increased at 20%, while the genes responsible for muscle development increased RNA abundance in MyoD at 20%, Mrf4 at 31%, PAX3 at 19%, and PAX7 at 16% observed in myotube development from myoblast when treated with NPC (P < 0.05). In NIH-3T3L1 cell, NPC treatment increased suppression on lipid accumulation at 40% (P < 0.05). In a follow-up in vivo investigation in a completely randomized design (Experiment 2), the same NPC tested in Experiment 1 at 0.53% dry matter (DM) did not affect intake of DM, whereas it tended to increase average daily gain compared with control without NPC supplementation (1.03 vs. 0.99; P = 0.08), leading to a tendency to increase gain-to-feed ratio (0.338 vs. 0.316; P = 0.12). While NPC supplementation did not influence hot carcass weight, animals treated with NPC decreased backfat thickness compared to those fed control (23.5 vs. 24.8 mm; P < 0.01). In summary, NPC from fruit improved growth performance and carcass trait due to their impacts on muscle cell development and fat deposition.

Synthesis and biological investigation of (+)-3-hydroxymethylartemisinin

Herein, we describe a biomimetic entry to (+)-3-hydroxymethylartemisinin (2) as well as to the artemisinin derivatives (+)-3-hydroxymethyl-9-desmethylartemisinin (16) and (+)-3-hydroxymethyl-9-epi-artemisinin (18), starting from the known and readily available chiral aldehyde 3 and alkyne 4. Subsequently, the synthesized compounds have been evaluated for their antimalarial activity against the drug-sensitive P. falciparum NF54 strain. All of them were inactive. In addition, they did not show any toxicity against L6 cells (a primary cell line derived from rat skeletal myoblasts). These results contribute to a better understanding of artemisinins mechanism of action.