The Ubiquitin/Proteasome System Mediates Entry and Endosomal Trafficking of Kaposi's Sarcoma-Associated Herpesvirus in Endothelial Cells

Ubiquitination, a post-translational modification, mediates diverse cellular functions including endocytic transport of molecules. Kaposi's sarcoma-associated herpesvirus (KSHV), an enveloped herpesvirus, enters endothelial cells primarily through clathrin-mediated endocytosis. Whether ubiquitination and proteasome activity regulates KSHV entry and endocytosis remains unknown. We showed that inhibition of proteasome activity reduced KSHV entry into endothelial cells and intracellular trafficking to nuclei, thus preventing KSHV infection of the cells. Three-dimensional (3-D) analyses revealed accumulation of KSHV particles in a cytoplasmic compartment identified as EEA1+ endosomal vesicles upon proteasome inhibition. KSHV particles are colocalized with ubiquitin-binding proteins epsin and eps15. Furthermore, ubiquitination mediates internalization of both KSHV and one of its receptors integrin β1. KSHV particles are colocalized with activated forms of the E3 ligase c-Cbl. Knock-down of c-Cbl or inhibition of its phosphorylation reduced viral entry and intracellular trafficking, resulting in decreased KSHV infectivity. These results demonstrate that ubiquitination mediates internalization of both KSHV and one of its cognate receptors integrin β1, and identify c-Cbl as a potential E3 ligase that facilitates this process

Genetic instability and anti-HPV immune response as drivers of infertility associated with HPV infection

Funding Information: RFBR grant 17–54-30002, Ministry of Science and Higher Education of the Russian Federation (Agreement No. 075–15–2019-1660) to Olga Smirnova. Publisher Copyright: © 2021, The Author(s).Human papillomavirus (HPV) is a sexually transmitted infection common among men and women of reproductive age worldwide. HPV viruses are associated with epithelial lesions and cancers. HPV infections have been shown to be significantly associated with many adverse effects in reproductive function. Infection with HPVs, specifically of high-oncogenic risk types (HR HPVs), affects different stages of human reproduction, resulting in a series of adverse outcomes: 1) reduction of male fertility (male infertility), characterized by qualitative and quantitative semen alterations; 2) impairment of couple fertility with increase of blastocyst apoptosis and reduction of endometrial implantation of trophoblastic cells; 3) defects of embryos and fetal development, with increase of spontaneous abortion and spontaneous preterm birth. The actual molecular mechanism(s) by which HPV infection is involved remain unclear. HPV-associated infertility as Janus, has two faces: one reflecting anti-HPV immunity, and the other, direct pathogenic effects of HPVs, specifically, of HR HPVs on the infected/HPV-replicating cells. Adverse effects observed for HR HPVs differ depending on the genotype of infecting virus, reflecting differential response of the host immune system as well as functional differences between HPVs and their individual proteins/antigens, including their ability to induce genetic instability/DNA damage. Review summarizes HPV involvement in all reproductive stages, evaluate the adverse role(s) played by HPVs, and identifies mechanisms of viral pathogenicity, common as well as specific for each stage of the reproduction process.publishersversionPeer reviewe

Abstract P2-11-13: The Effect of Positive Axillary Lymph Nodes on Symptoms, Physical Impairments, and Function

Abstract Purpose/Objective(s): The role of axillary lymph node (ALN) sampling in breast cancer (BC) treatment continues to evolve, and BC patients are recommended for post-operative regional nodal radiation therapy (RNRT) based on the number of positive ALN. RNRT is recommended for patients with 4 or more positive ALN, but it remains controversial in patients with 1–3 positive ALN and is rarely recommended for patients without positive ALN. Consideration of anticipated functional impairments often guides decision making. The purpose of this analysis is to investigate functional impairments in BC patients with varying numbers of positive ALN. Materials/Methods: 166 women were diagnosed with BC between 2001–05 and enrolled and treated in a prospective surveillance physical therapy program. 110 had zero positive ALN, 37 had 1–3 positive ALN, and 19 had 4 or more positive ALN on either sentinel LN biopsy or ALN dissection. Participants' upper extremity (UE) range of motion, strength, and limb volume were assessed pre-operatively and at 1, 3, 6, 9, and 12+ months post-operatively by a physical therapist. Limb volume was assessed using infrared optoelectronic perometry. At 12+ months, overall health status, UE symptoms and function, and physical activity levels were reported using standardized questionnaires. Chi-square tests and one-way ANOVA analyses were used to determine significance between groups (p ≤ 0.05). Results: Of these 166 patients, 94 received mastectomy and 72 received lumpectomy, while 41 received RNRT and 58 received whole breast tangent RT. No significant differences were found between groups with regard to age or race. The number of dissected LN was not significantly different between those patients with 1–3 positive ALN and 4 or more positive ALN. Rates of lymphedema and seroma were not significantly different between those patients with zero positive ALN and 1–3 positive ALN, and rates of cording were not significantly different between any of the groups. Increased lymphedema (p = 0.03) and seroma (p = 0.005) were seen in those patients with 4 or more positive ALN compared to those patients with zero positive ALN, but this may also be related to a significantly greater number of dissected LN in the former group. By 12+ months post-operatively, there were no differences in shoulder abduction, shoulder flexion, internal rotation, or external rotation between groups. No differences were seen between groups in self-reported fatigue, UE swelling or weakness, arm stiffness, or ability to climb stairs. Conclusions: Functional impairments represent an important category of morbidity for BC survivors and should be considered in pre-treatment decision making. The number of positive ALN may not correlate with increased impairment over the first year of treatment when a prospective surveillance physical therapy program is part of the plan of care. Additional research is needed to assess longer-term changes and the impact of axillary surgery and/or radiation in the context of aggregate effects of other BC treatment modalities. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-11-13.</jats:p

PD02-03: The Effect of Breast Conservation Therapy vs Mastectomy on Symptoms, Physical Impairments, and Function.

Abstract Background: Early-stage breast cancer (BC) patients choose between breast conservation therapy (BCT) and mastectomy based on comparable recurrence rates and overall survival. In the absence of mortality benefit, consideration of anticipated functional impairments could guide decision making. Although BCT offers less extensive surgery, the administration of radiation therapy (RT) may adversely impact upper extremity (UE) function. The purpose of this analysis is to investigate the effect of BCT vs modified radical mastectomy (MRM) without RT on functional impairments among BC survivors. Materials and Methods: 196 women diagnosed with BC between 200105 were enrolled and treated in a prospective surveillance physical therapy program. 115 received either BCT, including lumpectomy and whole breast RT, or MRM without RT and were analyzed for this report. Participants’ UE range of motion (ROM), strength, and limb volume were assessed pre-operatively and at 1, 3, 6, 9, and 12+ months post-operatively by a physical therapist. Limb volume was assessed using infrared optoelectronic perometry. At 12+ months, overall health status, UE symptoms and function, and physical activity levels were reported using standardized questionnaires. Analysis of variance estimated differences in impairments and self-reported symptoms and function. One-way ANOVA analysis was used to determine significance between groups (p ≤ 0.05). Results: 65 women (57.5%) received BCT and 50 women (42.5%) received MRM. No significant differences in age, BMI, stage, ER/PR status, and number of dissected lymph nodes were found between groups. At 1 month post-operatively, shoulder internal rotation (p=0.03), abduction (p=0.01), and flexion (p=0.004) were worse in post-MRM patients, with a trend towards worse external rotation (p=0.06). A higher rate of axillary cording was seen in patients post-MRM (p=0.02). By 12+ months post-operatively, there were no differences in any of the shoulder ROM variables. BCT patients reported, however, greater weakness (p=0.03) and diminished ability to perform heavy household tasks (p=0.03). There was no significant difference between BCT vs. MRM in rates of early lymphedema (40% vs 38%) or seroma (14% vs 22%). Conclusion: Functional impairments represent an important category of morbidity for BC survivors and should be considered in pre-treatment decision making. Post-operative RT as part of BCT may not contribute significantly to impairment over the first year of treatment. The presence of self-reported weakness and difficulty performing heavy household tasks at 12+ months suggest possible future functional deficits, especially considering the potentially progressive nature of RT-associated tissue changes. Additional research is needed to assess longer-term changes and the impact of RT in the context of aggregate effects of other BC treatment modalities. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD02-03.</jats:p

Abstract P2-11-14: Symptoms, Physical Impairments, and Function in Breast Cancer Patients with Negative Axillary Lymph Nodes

Abstract Purpose/Objective(s): Breast cancer (BC) patients are recommended for post-operative regional nodal radiation therapy (RNRT) based on the number of positive axillary lymph nodes (LN). While RNRT is recommended for patients with 4 or more positive LN, it remains controversial in patients with 1–3 positive LN. For these patients, consideration of anticipated functional impairments often guides decision making, but these considerations are confounded by the inseparable effects of disease in and treatment of the axilla. The purpose of this analysis is to investigate the effect of various therapies on functional impairments in BC patients without axillary disease. Materials/Methods: 166 women were diagnosed with BC between 2001–05 and enrolled and treated in a prospective surveillance physical therapy program. 110 had zero positive axillary LN on either sentinel LN biopsy or axillary LN dissection and were analyzed for this report. Participants' upper extremity (UE) range of motion, strength, and limb volume were assessed pre-operatively and at 1, 3, 6, 9, and 12+ months post-operatively by a physical therapist. Limb volume was assessed using infrared optoelectronic perometry. At 12+ months, overall health status, UE symptoms and function, and physical activity levels were reported using standardized questionnaires. Chi-square tests and one-way ANOVA analyses were used to determine significance between groups (p ≤ 0.05). Results: Of these 110 patients, 34 received mastectomy without RT, 21 received mastectomy with RNRT, 10 received lumpectomy alone, and 45 received lumpectomy with whole breast tangent RT. No significant differences were found between groups with regard to stage, ER/PR status, and number of dissected LN. Rates of lymphedema, cording, and seroma were not significantly different between groups. By 12+ months post-operatively, there were no differences in shoulder abduction, shoulder flexion, internal rotation, or external rotation between groups. No differences were seen between groups in self-reported fatigue, UE swelling or weakness, arm stiffness, or ability to climb stairs. Conclusions: Functional impairments represent an important category of morbidity for BC survivors and should be considered in pre-treatment decision making. In patients without axillary disease, post-operative RNRT or whole breast tangent RT may not contribute significantly to impairment over the first year of treatment when a prospective surveillance physical therapy program is part of the plan of care. Additional research is needed to assess longer-term changes and the impact of radiation in the context of the aggregate effect of disease burden combined with other BC treatment modalities. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-11-14.</jats:p

Making the Brain Glow: In Vivo Bioluminescence Imaging to Study Neurodegeneration

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