Blood Circulation-Prolonging Peptides for Engineered Nanoparticles Identified via Phage Display.

Sustaining blood retention for theranostic nanoparticles is a big challenge. Various approaches have been attempted and have demonstrated some success but limitations remain. We hypothesized that peptides capable of increasing blood residence time for M13 bacteriophage, a rod-shaped nanoparticle self-assembled from proteins and nucleic acids, should also prolong blood circulation for engineered nanoparticles. Here we demonstrate the feasibility of this approach by identifying a series of blood circulation-prolonging (BCP) peptides through in vivo screening of an M13 peptide phage display library. Intriguingly, the majority of the identified BCP peptides contained an arginine-glycine-aspartic acid (RGD) motif, which was necessary but insufficient for the circulation-prolonging activity. We further demonstrated that the RGD-mediated specific binding to platelets was primarily responsible for the enhanced blood retention of BCP1. The utility of the BCP1 peptide was demonstrated by fusion of the peptide to human heavy-chain ferritin (HFn), leading to significantly improved pharmacokinetic profile, enhanced tumor cell uptake and optimum anticancer efficacy for doxorubicin encapsulated in the HFn nanocage. Our results provided a proof-of-concept for an innovative yet simple strategy, which utilizes phage display to discover novel peptides with the capability of substantially prolonging blood circulation for engineered theranostic nanoparticles.

Executive Function Alternations of Breast Cancer Patients After Chemotherapy: Evidence From Resting-state Functional MRI.

RATIONALE AND OBJECTIVES: Chemotherapy has many side effects on breast cancer patients, including cognition and other brain functions impairment, which can be studied using functional magnetic resonance imaging (fMRI). Our study aimed at investigating the executive function alternations of breast cancer patients after chemotherapy using resting-state fMRI. MATERIALS AND METHODS: This study included 32 breast cancer patients (BC group) and 24 control subjects (HC group). The functional connectivity of the dorsolateral prefrontal cortex (DLPFC) of the two groups was calculated from the resting-state fMRI data, and the correlation between the strength of the right DLPFC's connectivity and the behavior performance was analyzed with two-tailed Pearson correlative analysis. RESULTS: Evaluation of the capability of processing various complex cognition events showed that the executive function of the BC group was impaired after chemotherapy in comparison with the HC group. The functional connectivities of the right DLPFC with the right inferior frontal gyrus, right medial frontal gyrus, and left superior temporal gyrus in the BC group were significantly decreased in comparison with those in the HC group, respectively. The executive deficits were found correlated with the functional connectivity between the right DLPFC and the right inferior frontal gyrus. Meantime, the functional connectivity from the right DLPFC to the right middle temporal gyrus and the precuneus was compensatorily increased in the BC group, respectively. CONCLUSIONS: These findings suggest that breast cancer patients after chemotherapy demonstrate executive control impairment, and provide evidence that the observed defects are correlated with alternations in the executive network of the brain.

Functional Connectivity Modulation by Acupuncture in Patients with Bell's Palsy.

Bell's palsy (BP), an acute unilateral facial paralysis, is frequently treated with acupuncture in many countries. However, the mechanism of treatment is not clear so far. In order to explore the potential mechanism, 22 healthy volunteers and 17 BP patients with different clinical duration were recruited. The resting-state functional magnetic resonance imaging scans were conducted before and after acupuncture at LI4 (Hegu), respectively. By comparing BP-induced functional connectivity (FC) changes with acupuncture-induced FC changes in the patients, the abnormal increased FC that could be reduced by acupuncture was selected. The FC strength of the selected FC at various stages was analyzed subsequently. Our results show that FC modulation of acupuncture is specific and consistent with the tendency of recovery. Therefore, we propose that FC modulation by acupuncture may be beneficial to recovery from the disease.

Long-term cognitive impairment of breast cancer patients after chemotherapy: A functional MRI study.

BACKGROUND: Chemotherapy, a prominent treatment for breast cancer (BC), can have detrimental side effects on the patient's cognitive functions including the executive function. However, the neurophysiological mechanism of the cognitive impairment remains unclear. OBJECTIVE: The purpose of this study is to explore long-term chemotherapy-related functional connectivity changes using fMRI and the relationship between the connectivity changes and the executive function impairment in breast cancer patients. METHODS: In this study, twenty-three breast cancer patients were treated with chemotherapy and twenty-six healthy subjects were recruited as the healthy control (HC) group. The functional connectivity of anterior cingulate cortex (ACC) was calculated from resting-state fMRI of the BC and control groups. The relationship between the functional connectivity of ACC and the executive function was further analyzed based on the patient' response time of the Stroop Interference Test. RESULTS: The results show that functional connectivity of ACC in the BC group is significantly lower than that in the control group. The correlation analysis within the BC group indicates that the functional connectivity of ACC was significantly correlated with the executive function. CONCLUSION: These findings provide evidence that the functional connectivity changes might be a pathophysiological basis for long-term chemotherapy-related cognitive dysfunction, along with executive function impairment in breast cancer patients.

The role of low levels of fullerene C60 nanocrystals on enhanced learning and memory of rats through persistent CaMKII activation.

Engineered nanomaterials are known to exhibit diverse and sometimes unexpected biological effects. Fullerene nanoparticles have been reported to specifically bind to and elicit persistent activation of hippocampal Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), a multimeric intracellular serine/threonine kinase central to Ca(2+) signal transduction and critical for synaptic plasticity, but the functional consequence of that modulation is unknown. Here we show that low doses of fullerene C60 nanocrystals (Nano C60), delivered through intrahippocampal infusion and without any obvious cytotoxicity in hippocampal neuronal cells, enhance the long-term potentiation (LTP) of rats. Intraperitoneal injection of 320 µg/kg of Nano C60, once daily for 10 days, also enhanced spatial memory of rats in addition to an increase of LTP. In parallel, both the IH and IP administration of Nano C60 increased the autonomous activity and the level of threonine 286 (T286) autophosphorylation of CaMKII, enhanced post-synaptic AMPA/NMDA ratio, and triggered time-dependent activation of ERK and CREB. Our results reveal a striking and highly unexpected ability of Nano C60 in positively modulating learning and memory, an effect that is most likely manifested through locking CaMKII in an active conformation, and may have significant implications for the potential therapeutic applications of fullerene C60, a classic engineered nanomaterial.

Nanoparticle as signaling protein mimic: robust structural and functional modulation of CaMKII upon specific binding to fullerene C60 nanocrystals.

In a biological environment, nanoparticles encounter and interact with thousands of proteins, forming a protein corona on the surface of the nanoparticles, but these interactions are oftentimes perceived as nonspecific protein adsorption, with protein unfolding and deactivation as the most likely consequences. The potential of a nanoparticle-protein interaction to mimic a protein-protein interaction in a cellular signaling process, characterized by stringent binding specificity and robust functional modulation for the interacting protein, has not been adequately demonstrated. Here, we show that water-suspended fullerene C60 nanocrystals (nano-C60) interact with and modulate the function of the Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), a multimeric intracellular serine/threonine kinase central to Ca(2+) signal transduction, in a fashion that rivals the well-documented interaction between the NMDA (N-methyl-d-aspartate) receptor subunit NR2B protein and CaMKII. The stable high-affinity binding of CaMKII to distinct sites on nano-C60, mediated by amino acid residues D246 and K250 within the catalytic domain of CaMKIIα, but not the nonspecific adsorption of CaMKII to diamond nanoparticles, leads to functional consequences reminiscent of the NR2B-CaMKII interaction, including generation of autonomous CaMKII activity after Ca(2+) withdrawal, calmodulin trapping and CaMKII translocation to postsynaptic sites. Our results underscore the critical importance of specific interactions between nanoparticles and cellular signaling proteins, and the ability of nano-C60 to sustain the autonomous kinase activity of CaMKII may have significant implications for both the biosafety and the potential therapeutic applications of fullerene C60.