Novel mechanism of drug resistance triggered by tumor-associated macrophages through Heat Shock Factor-1 activation.

Macrophages constitute a major part of tumor microenvironment, and most of existing data demonstrate their ruling role in the development of anti-drug resistance of cancer cell. One of the most powerful protection system is based on heat shock proteins whose synthesis is triggered by activated Heat Shock Factor-1 (HSF1); the inhibition of the HSF1 with CL-43 sensitized A549 lung cancer cells to the anti-cancer effect of etoposide. Notably, analyzing A549 tumor xenografts in mice we observed nest-like pattern of co-localization of A549 cells demonstrating enhanced expression of HSF1 with macrophages, and decided to check whether the above arrangement has a functional value for both cell types. It was found that the incubation of A549 or DLD1 colon cancer cells with either human monocytes or THP1 monocyte-like cells activated HSF1 and increased resistance to etoposide. Importantly, the same effect was shown when primary cultures of colon tumors were incubated with THP1 cells or with human monocytes. To prove that HSF1 is implicated in enhanced resistance caused by monocytic cells, we generated an A549 cell subline devoid of HSF1 which did not respond to incubation with THP1 cells. The pharmacological inhibition of HSF1 with CL-43 also abolished the effect of THP1 cells on primary tumor cells, highlighting a new target of tumor-associated macrophages in a cell proteostasis mechanism.

A non-randomized comparative study of olfactory and gustatory functions in children who recovered from COVID-19 (1-year follow-up).

The experimental group included 68 children over 6 years of age who had recovered from COVID-19. The control group included 22 children over 6 years of age who have never had COVID-19. Research methods included neurological examination, verification of cognitive status, examination by an otolaryngologist, and smell and taste assessment. The examination was performed 6-8 weeks after COVID-19 recovery and after 1 year in some patients. Children who recovered from COVID-19 had a reduction in their ability to smell compared to children who had never had COVID-19. The olfactory thresholds and taste identification scores after recovery from COVID-19 were identical, whether the parents had reported anosmia in their children during COVID-19 or not, and irrespective of hyperthermia level and the presence or absence of headache and hyperhidrosis during COVID-19. Analysis of correlation with neuropsychiatric symptoms showed no differences in the olfactory thresholds in children irrespective of the presence of neuropsychiatric symptoms (tics, tremors, enuresis, compulsive movements, seizures, speech disorders, attention deficit, and easy fatigability) both in general, and in particular among subjects performing or not any compulsive movements, and experiencing or not a combination of easy fatigability and daytime sleepiness. Evidence suggests that in children and adolescents, partial hyposmia is associated with depressive symptoms, varying in severity from low to high, but symptoms of depression were not caused by COVID-19 infection itself. Analysis in subgroups with different degrees of state and trait anxiety did not reveal any significant differences in the olfactory threshold. A re-examination of 21 children was performed after 1 year. An objective olfactometric examination showed that the sensitivity to odorants increased significantly. In 1 year, we compared the thresholds of smell in children who had COVID-19 and those who did not have this disease: olfactory sensitivity after COVID-19 in children is restored to normal values. Schulte correction test showed that none of 14 children with asthenic manifestations in the form of fluctuations or exhaustion when performing the test immediately after COVID-19 had these manifestations after 1 year. Thus, asthenization of cognitive activity was recorded within the next 1.5 months after suffering from COVID-19 but was absent after 1 year.

Combined Cytotoxic Effect of Inhibitors of Proteostasis on Human Colon Cancer Cells.

Despite significant progress in the diagnosis and treatment of colorectal cancer, drug resistance continues to be a major limitation of therapy. In this regard, studies aimed at creating combination therapy are gaining popularity. One of the most promising adjuvants are inhibitors of the proteostasis system, chaperone machinery, and autophagy. The main HSP regulator, HSF1, is overactivated in cancer cells and autophagy sustains the survival of malignant cells. In this work, we focused on the selection of combination therapy for the treatment of rectal cancer cells obtained from patients after tumor biopsy without prior treatment. We characterized the migration, proliferation, and chaperone status in the resulting lines and also found them to be resistant to a number of drugs widely used in the clinic. However, these cells were sensitive to the autophagy inhibitor, chloroquine. For combination therapy, we used an HSF1 activity inhibitor discovered earlier in our laboratory, the cardenolide CL-43, which has already been proven as an auxiliary component of combined therapy in established cell lines. CL-43 effectively suppressed HSF1 activity and Hsp70 expression in all investigated cells. We tested the autophagy inhibitor, chloroquine, in combination with CL-43. Our results indicate that the use of an inhibitor of HSF1 activity in combination with an autophagy inhibitor results in effective cancer cell death, therefore, this therapeutic approach may be a promising treatment regimen for certain patients.

Prevention of High Glucose-Mediated EMT by Inhibition of Hsp70 Chaperone.

Hyperglycemia may contribute to the progression of carcinomas by triggering epithelial-to-mesenchymal transition (EMT). Some proteostasis systems are involved in metastasis; in this paper, we sought to explore the mechanism of Hsp70 chaperone in EMT. We showed that knockdown of Hsp70 reduced cell migration capacity concomitantly with levels of mRNA of the Slug, Snail, and Twist markers of EMT, in colon cancer cells incubated in high glucose medium. Conversely, treatment of cells with Hsp70 inducer U-133 were found to elevate cell motility, along with the other EMT markers. To prove that inhibiting Hsp70 may reduce EMT efficiency, we treated cells with a CL-43 inhibitor of the HSF1 transcription factor, which lowered Hsp70 and HSF1 content in the control and induced EMT in carcinoma cells. Importantly, CL-43 reduced migration capacity, EMT-linked transcription factors, and increased content of epithelial marker E-cadherin in colon cancer cells of three lines, including one derived from a clinical sample. To prove that Hsp70 chaperone should be targeted when inhibiting the EMT pathway, we treated cancer cells with 2-phenylethynesulfonamide (PES) and demonstrated that the compound inhibited substrate-binding capacity of Hsp70. Furthermore, PES suppressed EMT features, cell motility, and expression of specific transcription factors. In conclusion, the Hsp70 chaperone machine efficiently protects mechanisms of the EMT, and the safe inhibitors of the chaperone are needed to hamper metastasis at its initial stage.

Calcium Administration During Weaning From Cardiopulmonary Bypass: A Narrative Literature Review.

The search for safe and effective patient management strategies during weaning from cardiopulmonary bypass is ongoing; intravenous calcium is occasionally used as a first-line drug. The physiologic role of calcium suggests that it can support the function of the cardiovascular system during this critical period. Patients may be mildly hypocalcemic after cardiopulmonary bypass; however, this degree of hypocalcemia does not significantly impair the cardiovascular system. The transient beneficial effects of calcium administration (increase in arterial blood pressure, systemic vascular resistance, cardiac index, stroke volume, and coronary perfusion pressure) might be helpful in cases of moderate contractility reduction or vasoplegia. Nonetheless, effects on clinically relevant endpoints are unknown, and possible systemic side effects, such as transient reduction in internal mammary artery graft flow, attenuation of the effects of ß-sympathomimetics, "stone heart" phenomenon, and pancreatic cellular injury, may limit the use of calcium salts. Further studies are needed to expand the understanding of the effects of calcium administration on patient outcomes.