Combining Stimulus-Triggered Release and Active Targeting Strategies Improves Cytotoxicity of Cytochrome c Nanoparticles in Tumor Cells.

Proteins often possess highly specific biological activities that make them potential therapeutics, but their physical and chemical instabilities during formulation, storage, and delivery have limited their medical use. Therefore, engineering of nanosized vehicles to stabilize protein therapeutics and to allow for targeted treatment of complex diseases, such as cancer, is of considerable interest. A micelle-like nanoparticle (NP) was designed for both, tumor targeting and stimulus-triggered release of the apoptotic protein cytochrome c (Cyt c). This system is composed of a Cyt c NP stabilized by a folate-receptor targeting amphiphilic copolymer (FA-PEG-PLGA) attached to Cyt c through a redox-sensitive bond. FA-PEG-PLGA-S-S-Cyt c NPs exhibited excellent stability under extracellular physiological conditions, whereas once in the intracellular reducing environment, Cyt c was released from the conjugate. Under the same conditions, the folate-decorated NP reduced folate receptor positive HeLa cell viability to 20%, while the same complex without FA only reduced it to 80%. Confocal microscopy showed that the FA-PEG-PLGA-S-S-Cyt c NPs were internalized by HeLa cells and were capable of endosomal escape. The specificity of the folate receptor-mediated internalization was confirmed by the lack of uptake by two folate receptor deficient cell lines: A549 and NIH-3T3. Finally, the potential as antitumor therapy of our folate-decorated Cyt c-based NPs was confirmed with an in vivo brain tumor model. In conclusion, we were able to create a stable, selective, and smart nanosized Cyt c delivery system.

Disparities in Hepatocellular Cancer Screening in Cirrhotic Patients With Psychiatric Disorders.

Background Patients with psychiatric disorders are at an increased risk of developing liver diseases, including hepatocellular carcinoma (HCC). HCC is a leading cause of cancer-related deaths in the United States. The aim of this study was to re-examine the association of psychiatric illness with HCC and assess its impact on screening practices and the outcomes of HCC. Materials and methods We performed a retrospective manual chart review of all patients diagnosed with HCC at a major safety-net hospital in Cleveland, Ohio, from January 2010 to December 2019. Patients were divided into two groups, those with and those without psychiatric illness. The patient characteristics recorded included psychiatric illnesses, etiology of liver disease, radiographic screening intervals, and tumor board recommendations upon initial diagnosis. We analyzed data using Statistical Product and Service Solutions version 26.0 (IBM Corp., Armonk, NY). We analyzed the qualitative and quantitative differences between the groups using the chi-square or Fisher's exact tests for categorical variables and t-test for continuous variables. Results There were a total of 393 patients with a diagnosis of HCC. Among them, 128 (32.5%) were diagnosed with at least one psychiatric illness. Fewer patients with psychiatric illness (33.6%) underwent screening within six months before being diagnosed with HCC compared to those without psychiatric illness (49.8%) (p = 0.002). Patients with psychiatric illness (71.1%) were more likely to have been seen by a gastroenterologist or hepatologist before their diagnosis of HCC compared to those without psychiatric illness (55.1%) (p =0.002). Patients with psychiatric illness were more likely to be offered systemic chemotherapy or hospice (39.1%) compared to those without psychiatric illness (29.1%) (p =0.039). Discussion A significant number of HCC patients in our study group have an underlying psychiatric illness. Patients with psychiatric disorders are prone to high-risk behaviors, likely predisposing them to chronic liver disease and HCC. Patients with psychiatric disorders are less compliant with screening practices. Our findings suggest that psychiatric illnesses tend to be diagnosed with more extensive HCC, which is less amenable to curative treatment. Significant efforts need to be made to identify barriers to HCC screening in cirrhotic patients with psychiatric disorders.

Targeted Delivery of Nanoparticulate Cytochrome C into Glioma Cells Through the Proton-Coupled Folate Transporter.

In this study, we identified the proton-coupled folate transporter (PCFT) as a route for targeted delivery of drugs to some gliomas. Using the techniques of confocal imaging, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and small interfering (siRNA) knockdown against the PCFT, we demonstrated that Gl261 and A172 glioma cells, but not U87 and primary cultured astrocytes, express the PCFT, which provides selective internalization of folic acid (FA)-conjugated cytochrome c-containing nanoparticles (FA-Cyt c NPs), followed by cell death. The FA-Cyt c NPs (100 µg/mL), had no cytotoxic effects in astrocytes but caused death in glioma cells, according to their level of expression of PCFT. Whole-cell patch clamp recording revealed FA-induced membrane currents in FA-Cyt c NPs-sensitive gliomas, that were reduced by siRNA PCFT knockdown in a similar manner as by application of FA-Cyt c NPs, indicating that the PCFT is a route for internalization of FA-conjugated NPs in these glioma cells. Analysis of human glioblastoma specimens revealed that at least 25% of glioblastomas express elevated level of either PCFT or folate receptor (FOLR1). We conclude that the PCFT provides a mechanism for targeted delivery of drugs to some gliomas as a starting point for the development of efficient methods for treating gliomas with high expression of PCFT and/or FOLR1.