scholarly journals Assessment of the effectiveness of the use of tumoroids for personalized drug therapies for solid tumors

2021 ◽  
Vol 67 (6) ◽  
pp. 815-828
Author(s):  
Anna Danilova ◽  
Tatiana Nekhaeva ◽  
Natalia Efremova ◽  
Aleksei Novik ◽  
Anton Zozulia ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Monolayer Culture ◽  
Osteogenic Sarcoma ◽  
Test System ◽  
Chemotherapeutic Agents ◽  
Model Systems ◽  
Predictive Test ◽  
Layer System ◽  
Medicine Research

The approach to the management of cancerous neoplasms, which aims to define the effective curative strategies in each patient, defines the requirement for the elaboration and use of modelling systems that replicate the structures and the biology of human solitary tumors. Three-dimensional cultures of spheroids/tumoroids, which are multi-cell aggregates of malignized cells, can create the intercellular connections of interest, gradients of the nutrients and oxygen, and cell polarity, all of which are absent in the conventional two-dimensional single-layer system. The present work is dedicated to a comparison study of in vitro viability and invasiveness of solid tumor cells of patients under the effect of chemopreparations and their combinations in view of evaluating the efficacy of the 3D-cell modelling system in the translational personalized medicine context. Cell cultures of patients who were treated at the N.N. Petrov National Medicine Research Center of oncology were used as a basis for the development of 3D-cell models. N.N. Petrov NMRC in 2015-2021. Tumor tissue pieces were acquired intraoperatively: 1 - leiomyosarcoma (LMS), 1 - rhabdomyosarcoma (RMS), 1 - synovial sarcoma (SS), 2 - myxofibrosarcoma (MFS), 2 - osteogenic sarcoma (OS), 1 - skin melanoma (MC), 1 - breast cancer (BC) (n=9). Our individual comparison of the effectiveness of in vitro chemotherapeutic agents against tumor cells of various origins cultivated in 2D and 3D model systems with real clinically relevant cases confirmed that the monolayer culture as the test system was less adequate for selecting and personalizing the treatment of malignant tumor patients: the 3D cell system proved itself in 77.7% of cases, and the monolayer culture - in 44.4% of cases. The combination of doxorubicin/iforsfamide and paclitaxel significantly suppressed the motility in the matrigel of spheroid cells, but did not affect tumor cell viability, which was seen in all but OS #921 and MK #929 cases. The cultivation of tumor cells in form of spheroids/tumoroids allows to utilize them as more adequate pre-clinical model as individual predictive test-system, enabling the personalized selection of therapy.  

scholarly journals Mitotic arrest affects clustering of tumor cells

2021 ◽  
Author(s):  
Julia Bonnet ◽  
Lise Rigal ◽  
Odile Mondesert ◽  
Renaud Morin ◽  
Gaelle Corsaut ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Cell ◽  
Cell Aggregation ◽  
Metastatic Potential ◽  
Mitotic Arrest ◽  
Chemotherapeutic Agents ◽  
The Impact ◽  
Mcf 7

Abstract Background Cancer cell aggregation is a key process involved in the formation of tumor cell clusters. It has recently been shown that clusters of circulating tumor cells (CTCs) have an increased metastatic potential compared to isolated circulating tumor cells. Several widely used chemotherapeutic agents that target the cytoskeleton microtubules and cause cell cycle arrest at mitosis have been reported to modulate CTC number or the size of CTC clusters. Results In this study, we investigated in vitro the impact of mitotic arrest on the ability of breast tumor cells to form clusters. By using live imaging and quantitative image analysis, we found that MCF-7 cancer cell aggregation is compromised upon incubation with paclitaxel or vinorelbine, two chemotherapeutic drugs that target microtubules. In line with these results, we observed that MCF-7 breast cancer cells experimentally synchronized and blocked in metaphase aggregated poorly and formed loose clusters. To monitor clustering at the single-cell scale, we next developed and validated an in vitro assay based on live video-microscopy and custom-designed micro-devices. The study of cluster formation from MCF-7 cells that express the fluorescent marker LifeAct-mCherry using this new assay allowed showing that substrate anchorage-independent clustering of MCF-7 cells was associated with the formation of actin-dependent highly dynamic cell protrusions. Metaphase-synchronized and blocked cells did not display such protrusions, and formed very loose clusters that failed to compact. Conclusions Altogether, our results suggest that mitotic arrest induced by microtubule-targeting anticancer drugs prevents cancer cell clustering and therefore, could reduce the metastatic potential of circulating tumor cells.

scholarly journals Prediction of in Vivo Cytotoxicity of Chemotherapeutic Agents by Their Effect on Malignant Leukocytes in Vitro

Blood ◽  
1967 ◽  
Vol 30 (2) ◽  
pp. 176-188 ◽  
Author(s):  
MARTIN J. CLINE
Keyword(s):  
Test System ◽  
Chemotherapeutic Agents ◽  
Response To Treatment ◽  
In Vitro Test ◽  
Malignant Cells ◽  
Vitro Test ◽  
In Vitro Effects ◽  
In Vivo Cytotoxicity

Abstract In order to develop a test system for predicting the response to chemotherapeutic agents, leukocytes from patients with leukemia and leukolymphosarcoma were cultured in vitro and the effect of several drugs on the incorporation of H3-uridine into ribonucleic acid was measured. Cortisol, vincristine and cytosine arabinoside at concentrations near the therapeutic range produced inhibition of H3-uridine incorporation in sensitive leukocytes. The in vitro effects of 6-mercaptopurine and methotrexate were variable. In 39 trials on 25 patients with leukemia or lymphosarcoma, the in vitro test was used successfully to predict the response to treatment with prednisone and vincristine. It was concluded that the in vitro test system can predict the in vivo cytotoxicity of certain drugs for malignant cells, although it cannot be used to predict the likelihood of the induction of remissions with these drugs.

Combining galiximab with the chemotherapeutic agents fludarabine or doxorubicin improves efficacy in animal models of lymphoma

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 3040-3040 ◽  
Author(s):  
H. K. Hariharan ◽  
T. Murphy ◽  
D. Clanton ◽  
L. Berquist ◽  
P. Chu ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Animal Models ◽  
Clinical Benefit ◽  
Therapeutic Potential ◽  
Xenograft Model ◽  
Chemotherapeutic Agents ◽  
Model Systems ◽  
Anti Cd20

3040 Background: Galiximab, a primatized monoclonal antibody that binds with high affinity to CD80 and mediates antibody- dependent, cell-mediated cytotoxicity in vitro, is currently under investigation for the treatment of follicular non-Hodgkin’s lymphoma (NHL). In a phase I/II monotherapy study, galiximab produced an overall response rate of 11%, and tumor reductions were observed in 46% of patients. Initial clinical trials also demonstrate that galiximab is well tolerated and suggest that combining galiximab with rituximab (anti-CD20) provides clinical benefit. These results are consistent with preclinical studies in murine lymphoma xenograft model systems, which demonstrate the superiority of combination therapy. Methods: To further define the therapeutic potential of galiximab, the Raji subcutaneous and the SKW disseminated lymphoma murine xenograft models were used to define the in vivo efficacy of galiximab alone or in combination with fludarabine or doxorubicin. Similar studies were performed with rituximab. Results: In the Raji model, both galiximab and rituximab exhibited maximal inhibition of the growth of preestablished (150-mg) tumors at a dose of 3 mg/kg/wk. Interestingly, higher doses of galiximab (but not rituximab) showed reduced inhibition. Galiximab (3 mg/kg/wk) inhibited tumor growth alone (P<0.0001 vs. control) and showed significantly enhanced activity when combined with fludarabine (50 or 100 mg/kg daily for 5 days; P<0.0002 vs. galiximab alone and P<0.003 vs. fludarabine alone). Similar results were observed with rituximab. In the SKW model, treatment with galiximab (5 mg/kg/wk for 6 doses) significantly enhanced survival compared with a control (P<0.0001) or doxorubicin (2.5 mg/kg/day for 3 doses; P<0.0001). Studies combining fludarabine or doxorubicin with both galiximab and rituximab are ongoing. Conclusions: Studies in animal models of lymphoma indicate that galiximab may provide clinical benefit when used in combination with chemotherapeutic agents such as fludarabine and doxorubicin, and provide a rationale for the investigation of these novel chemoimmunotherapy combinations in clinical trials. No significant financial relationships to disclose.

Proliferation, Esterase Activity, and Propidium Iodide Exclusion in Urologic Tumor Cells After in Vitro Exposure to Chemotherapeutic Agents

1986 ◽  
Vol 135 (5) ◽  
pp. 1091-1100 ◽  
Author(s):  
Robert C. Flanigan ◽  
Edward J. Pavlik ◽  
John R. van Nagell ◽  
Kathryn Keaton ◽  
Daniel E. Kenady
Keyword(s):  
Tumor Cells ◽  
Propidium Iodide ◽  
Esterase Activity ◽  
Chemotherapeutic Agents ◽  
In Vitro Exposure

Nongenotoxic activation of the p53 pathway as a therapeutic strategy for multiple myeloma

Blood ◽  
2005 ◽  
Vol 106 (10) ◽  
pp. 3609-3617 ◽  
Author(s):  
Thorsten Stühmer ◽  
Manik Chatterjee ◽  
Martin Hildebrandt ◽  
Pia Herrmann ◽  
Hella Gollasch ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Tumor Cells ◽  
Apoptotic Cell ◽  
Rare Event ◽  
Chemotherapeutic Agents ◽  
In Vitro Toxicity ◽  
P53 Pathway ◽  
Myeloma Cells ◽  
P53 Signaling

AbstractMutation of p53 is a rare event in multiple myeloma, but it is unknown if p53 signaling is functional in myeloma cells, and if targeted nongenotoxic activation of the p53 pathway is sufficient to kill tumor cells. Here, we demonstrate that treatment of primary tumor samples with a small-molecule inhibitor of the p53–murine double minute 2 (MDM2) interaction increases the level of p53 and induces p53 targets and apoptotic cell death. Significantly, given the importance of the bone marrow microenvironment for the support and drug resistance of myeloma cells, tumor cells undergo effective apoptosis also in the presence of stromal cells, which themselves appear to tolerate exposure to nutlin-3. The in vitro toxicity of nutlin-3 was similar to that of the genotoxic drug melphalan. Because nutlin-mediated p53 activation is not dependent on DNA damage, MDM2 antagonists may help to avoid or reduce the severe genotoxic side effects of chemotherapeutic agents currently used to treat multiple myeloma. Therefore, MDM2 antagonists may offer a new treatment option for this disease.

scholarly journals RBE of "Prometeus" Facility Protons for Irradiation of Tumor Cells in vitro with One and Three Fields

2018 ◽  
Vol 3 (3) ◽  
pp. 309
Author(s):  
Nasedkina N.V. ◽  
Beketov E.E. ◽  
Isaeva E.V. ◽  
Malakhov E.P. ◽  
Golovanova O.Y. ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Bragg Peak ◽  
Dose Interval ◽  
Test System ◽  
Cell Irradiation ◽  
Biological Effectiveness ◽  
System Cell ◽  
Single Field ◽  
Russian Medical

The study was aimed to the biological effectiveness of the proton scanning beam of the first Russian medical facility. The clonogenic assay of B-16 tumor cells was used as a test system. Cell irradiation was carried out in a suspension condition in a water phantom. Single and three-field exposures were studied. The dose interval was 2-8 Gy. The energy range from 47.5 to 92.0 MeV was used for the Bragg peak formation. The relative biological effectiveness of protons comparing to gamma-rays was 1.2 for single-field and 1.5 for three-field irradiation. The results obtained agree with literature data related to the used cell culture (B-16) and linear energy transfer range (3÷8 keV/µm).

3D культура меланоцитов как тест-система и клеточная модель для изучения патологии меланогенеза

2018 ◽  
pp. 265-268
Author(s):  
И.Н. Сабурина ◽  
Е.В. Джуссоева ◽  
А.А. Горкун ◽  
И.М. Зурина ◽  
Н.В. Кошелева ◽  
...  
Keyword(s):  
Animal Models ◽  
Functional Activity ◽  
Monolayer Culture ◽  
Skin Pigmentation ◽  
Test System ◽  
Specific Cell ◽  
Protective Properties ◽  
3D Cultivation

Функциональная активность меланоцитов обусловливает защитные свойства кожи против воздействия ультрафиолета, вызывающего фотостарение. Однако изучение меланоцитов in vivo затруднено тем, что необходимо использовать животные модели и трудоемкие методы анализа, а культивирование клеток в монослойной культуре in vitro сопряжено с потерей тканеспецифичных маркеров клеток. Данная работа посвящена получению и изучению сфероидов из меланоцитов, так как 3D культивирование меланоцитов в виде сфероидов может сохранить их фенотип и функциональность. Исследование проводили на первичной культуре меланоцитов кожи человека. Клетки культивировали в монослое в полной ростовой среде до 4 пассажа. Далее клетки помещали на агарозные планшеты с микролунками в посевной плотности 3,3 х 10 кл./мл. Анализ полученных сфероидов производили с помощью фотометрии, иммуноцитохимии и ПЦР в реальном времени. Было показано, что, при культивировании в монослое, к 4 пассажу снижалось количество синтезируемого меланина. Тогда как в 3D условиях меланоциты формировали компактные сфероиды, внутри которых в процессе культивирования не только сохранялся синтез, но и накапливался меланин. Была выявлена экспрессия специфических генов TYR и MCR1, и увеличивался синтез белков, участвующих в меланогенезе - gp100 и MITF. Таким образом, в данном исследовании было показано, что меланоциты in vitro способны формировать длительно живущие, жизнеспособные 3D структуры - сфероиды, с сохранением фенотипа и синтеза тканеспецифичных маркеров. Поэтому данные сфероиды могут быть успешно использованы как тест-системы для оценки эффективности препаратов, направленных на регуляцию уровня пигментации кожи. The functional activity of melanocytes determines protective properties of the skin against the effect of ultraviolet radiation, which causes photo-aging. However, studying melanocytes in vivo and in an in vitro monolayer culture is difficult because animal models and laborious analytical methods are required, and the culturing is associated with loss of tissue-specific cell markers. Since 3D cultivation of melanocytes in the form of spheroids can preserve their phenotype and functionality, this work focused on obtaining and studying melanocyte spheroids. The study was conducted using a primary culture of human skin melanocytes. Cells were cultured in a monolayer to the fourth passage. Then these cells were placed in agarose plates with microwells at the cell suspension concentration of 3.3 х 10 in vitro . These features make the melanocyte spheroids a convenient test-system for studying toxicity and efficiency of drugs targeted at regulation of skin pigmentation.

scholarly journals Insulin enhancement of the antitumor activity of chemotherapeutic agents in colorectal cancer is linked with downregulating PIK3CA and GRB2

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Siddarth Agrawal ◽  
Marta Woźniak ◽  
Mateusz Łuc ◽  
Sebastian Makuch ◽  
Ewa Pielka ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Western Blotting ◽  
Expression Patterns ◽  
Critical Role ◽  
Chemotherapeutic Agents ◽  
Significant Inhibition

Abstract The present state of cancer chemotherapy is unsatisfactory. New anticancer drugs that marginally improve the survival of patients continue to be developed at an unsustainably high cost. The study aimed to elucidate the effects of insulin (INS), an inexpensive drug with a convincing safety profile, on the susceptibility of colon cancer to chemotherapeutic agents: 5-fluorouracil (FU), oxaliplatin (OXA), irinotecan (IRI), cyclophosphamide (CPA) and docetaxel (DOC). To examine the effects of insulin on cell viability and apoptosis, we performed an in vitro analysis on colon cancer cell lines Caco-2 and SW480. To verify the results, we performed in vivo analysis on mice bearing MC38 colon tumors. To assess the underlying mechanism of the therapy, we examined the mRNA expression of pathways related to the signaling downstream of insulin receptors (INSR). Moreover, we performed Western blotting to confirm expression patterns derived from the genetic analysis. For the quantification of circulating tumor cells in the peripheral blood, we used the maintrac method. The results of our study show that insulin-pretreated colon cancer cells are significantly more susceptible to commonly used chemotherapeutics. The apoptosis ratio was also enhanced when INS was administered complementary to the examined drugs. The in vivo study showed that the combination of INS and FU resulted in significant inhibition of tumor growth and reduction of the number of circulating tumor cells. This combination caused a significant downregulation of the key signaling substrates downstream of INSR. The results indicate that the downregulation of PIK3CA (phosphatidylinositol 3-kinase catalytic subunit alpha), which plays a critical role in cell signaling and GRB2 (growth factor receptor-bound protein 2), a regulator of cell proliferation and differentiation may be responsible for the sensitizing effect of INS. These findings were confirmed at protein levels by Western blotting. In conclusion, these results suggest that INS might be potentially applied to clinical use to enhance the therapeutic effectiveness of chemotherapeutic drugs. The findings may become a platform for the future development of new and inexpensive strategies for the clinical chemotherapy of tumors.

Copper-Complexed Melphalan (MOC*M), a Member of a New Class of Anti-Leukemic and Anti-Tumouric Substances for Treatment of Primary and Relapsed Acute Lymphoblastic Leukemia in Childhood.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4457-4457
Author(s):  
Aram Prokop ◽  
Corazon Frias ◽  
Guenter Henze ◽  
Swetlana Sadolinnaya ◽  
Valeriy Tatarskiy
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Tumor Cells ◽  
Dna Fragmentation ◽  
Lymphoblastic Leukemia ◽  
Model Systems ◽  
Cytostatic Drugs ◽  
Cytostatic Agents ◽  
New Class

Abstract Acute lymphoblastic leukemia (ALL) is the most frequent malignant disease of childhood. Despite a relatively good prognosis (survival 80 %), approximately ¼ of the patients suffer from relapses with a much poorer prognosis (survival 40 %). If a complete remission through chemotherapy is not achieved, the patients will not survive. Thus, the search for new cytostatic substances which can break the resistance against conventional cytostatic drugs is of great interest. We developed a new class of copper-containing cytostatic agents with apoptosis-inducing properties. The present study deals with 20 children, who suffer from de novo ALL or relapsed ALL. In vitro measurement of DNA-fragmentation in primary lymphoblasts of the children showed, that the copper-complexed cytostatic drugs are considerably more effective, compared to conventional analogues and other cytostatic drugs (cytarabine p<0.002, vincristine p< 0.006) used against childhood ALL. Furthermore, the new copper-containing analogues overcome drug resistance against doxorubicin (p<0.001) in vitro. In addition, the prototype of copper-complexed drug analogues, MOC*M, a melphalan-copper-acetoacetonate-complex, has synergistic effects in apoptosis induction combined with melphalan or conventional drugs in therapy of ALL in childhood like vincristin, doxorubicin and cytarabine. Experiments revealed that MOC*M specifically induces apoptosis, as evidenced by DNA fragmentation and dissipation of the mitochondrial membrane potential. MOC*M induces cell death, which was functionally characterized by the use of different cellular model systems being devoid of defined molecular parts of the apoptosis machinery. MOC*M triggers apoptosis in a Bcl-2-independent manner in the multi-resistant melanoma cell line MelHO with a 30-fold over-expression of Bcl-2. In vitro and in vivo experiments on mice with tumors sarcom S-180, melanoma B-16 and adenocarcenom in the large intestine proved a high anti-tumor activity of MOC*M with anti-metastasis and immunizing properties without any side effects in kidney or liver. Thus, MOC*M is able to prolong the life of animals with leucosis L-1210 and P-388. We could show that the accumulation of the tritium-labelled MOC*M compounds took place mainly in the tumor cells in vivo. Moreover, MOC*M is also inhibiting glycolysis in the tumor cells. The result of pre-clinical tests with MOC*M preparations, tested on a limited quota of oncological patients with different tumors, was a very large spectrum of anti-tumor and anti-leukemic activities. Further MOC*M has an immense tolerability in vivo. All in all, copper-containing cytostatic drugs comprise an innovative, highly promising class of cytostatic agents for cancer and leukemia therapy, especially for the therapy of relapsed ALL in childhood.

Effect of lenalidomide on the antiproliferative effect of gemcitabine against pancreatic tumor cells and on immune-mediated pancreatic cancer cell death

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e14635-e14635
Author(s):  
L. Wu ◽  
W. Liu ◽  
C. Galustian ◽  
P. Schafer ◽  
A. G. Dalgleish ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cell Death ◽  
Tumor Cells ◽  
Solid Tumor ◽  
Proliferative Activity ◽  
Pancreatic Tumor ◽  
Pancreatic Cancer Cell ◽  
Chemotherapeutic Agents ◽  
Immune Mediated

e14635 Background: Lenalidomide is an immunomodulatory and anti-angiogenic agent that has demonstrated activity against a range of hematological malignancies. Despite evidence of direct anti-proliferative activity against hematological cells in vitro, there is no evidence of single agent direct activity against solid tumor cells in vitro. To take advantage of its known immune-enhancing properties alongside direct anti-tumor agents, lenalidomide is being advanced in solid tumor indications in combination with other agents. There are few data regarding the combination of lenalidomide and standard of care chemotherapeutic agents, such as gemcitabine. Methods: Here, we assess the effects of lenalidomide alone, and in combination with gemcitabine, on pancreatic cancer cell growth and survival, and the ability of lenalidomide to enhance the ability of human PBMC to kill allogeneic pancreatic tumor cells (BxPC3, PANC-1 and MiaPaCa) in a PBMC:tumor cell co-culture model. Results: Lenalidomide alone had no effect on the proliferation of pancreatic cancer cells (BxPC-3 and Panc-1) whereas gemcitabine had moderate anti-proliferative activity. With combination therapy there was clear synergistic enhancement of anti-proliferative activity in both cell lines and additive effects were observed in a BxPC-3 xenograft mouse model of pancreatic cancer. About 20% of tumor cells were sensitive to immune-mediated cell death and, for BxPC3, this was increased significantly in the presence of lenalidomide. Lenalidomide significantly and dose-dependently enhanced immune-mediated killing (both T and NK cells are required for tumor cell killing in this model). For PANC-1 and MiaPaCa, immune-mediated killing was also increased by lenalidomide, albeit non-significantly. Conclusions: These results suggest that, in addition to anti-angiogenic and other effects within the tumor microenvironment, lenalidomide may act as an immune adjuvant to enhance the recognition and apoptosis of tumor cells by host T and NK cells. These studies support the potential utility of lenalidomide in combination with chemotherapeutic agents, gemcitabine in particular, in the treatment of patients with solid tumors including pancreatic cancer. [Table: see text]

Sign in / Sign up

Export Citation Format

Share Document