Understanding the Role of Eed Deletion in Medulloblastoma

Medulloblastoma, the most common malignant pediatric brain cancer, is uniquely sensitive to DNA damage-inducing therapies, with conventional treatment resulting in an 80% 5-year survival rate. Researching the apoptotic pathways that make treatment effective in some tumors and how those pathways contribute to resistance in others may identify therapies that reduce the need for toxic radiation and chemotherapy. In cerebellar development, physiologic Sonic Hedgehog (SHH) signaling drives proliferation of cerebellar granule neuron progenitors (CGNPs). Similarly, pathologic SHH hyperactivation drives proliferation in SHH-subgroup medulloblastoma, which makes up 30% of medulloblastoma cases. SHH signaling upregulates target genes in part by preventing H3K27 trimethylation marks via the JMJD3/KDM6B demethylase complex. In differentiated cerebellar neurons, where SHH signaling is low, the polycomb repressive complex 2 (PRC2) silences SHH target genes by trimethylating H3K27 residues in regulatory regions. Our data show that blocking the PRC2 through genetic deletion has varying effects in different contexts. When the PRC2-component Eed is deleted in SHH-subtype medulloblastomas that form in mice engineered for SHH hyperactivation, the tumors show initially slower growth compared to tumors in Eed-intact controls; however, the mice have significantly poorer survival. Therefore, this study aims to understand the mechanisms of initially reduced tumor growth and ultimately more rapid tumor progression. Our recent single cell gene expression analysis showed that Eed deletion in CGNPs induced genes involved in muscle cell differentiation, including myogenin and troponin. We will characterize Eed-deleted medulloblastoma using fluorescence-activated cell sorting (FACS) as well as immunohistochemistry (IHC). The first aim of this study is to elucidate how cell cycle dynamics are altered in Eed-deleted medulloblastomas. The second aim of the study tests whether Eed deletion alters the levels of cell cycle markers, cell death, and aberrant expression of muscular proteins in SHH-driven medulloblastoma. Our cell cycle studies show that Eed deletion slows tumor growth in early development, but the tumor later develops EED independence, resulting in more rapid progression and decreased survival. Additionally, our data show that Eed deletion alters the expression of proteins related to proliferation, differentiation, apoptosis, and muscle development.Bachelor of Scienc

Assessment of the Field Utility of a Rapid Point-of-Care Test for SARS-CoV-2 Antibodies in a Household Cohort.

Point-of-care (POC) tests to detect SARS-CoV-2 antibodies offer quick assessment of serostatus after natural infection or vaccination. We compared the field performance of the BioMedomics COVID-19 IgM/IgG Rapid Antibody Test against an ELISA in 303 participants enrolled in a SARS-CoV-2 household cohort study. The rapid antibody test was easily implemented with consistent interpretation across 14 users in a variety of field settings. Compared with ELISA, detection of seroconversion lagged by 5 to 10 days. However, it retained a sensitivity of 90% (160/177, 95% confidence interval [CI] 85-94%) and specificity of 100% (43/43, 95% CI 92-100%) for those tested 3 to 5 weeks after symptom onset. Sensitivity was diminished among those with asymptomatic infection (74% [14/19], 95% CI 49-91%) and early in infection (45% [29/64], 95% CI 33-58%). When used appropriately, rapid antibody tests offer a convenient way to detect symptomatic infections during convalescence

Household transmission of SARS-CoV-2 in the United States: living density, viral load, and disproportionate impact on communities of color

Households are hotspots for SARS-CoV-2 transmission. In the US, the COVID-19 pandemic has had a disproportionate impact on communities of color. Between April-October 2020, the CO-HOST prospective cohort study enrolled 100 COVID-19 cases and 208 of their household members in North Carolina, including 44% who identified as Hispanic or non-white. Households were enrolled a median of 6 days from symptom onset in the index case. Incident secondary cases within the household were detected by quantitative PCR of weekly nasal swabs (days 7, 14, 21) or by seroconversion at day 28.Excluding 73 household contacts who were PCR-positive at baseline, the secondary attack rate among household contacts was 32% (33/103, 95% CI 22%-44%). The majority of cases occurred by day 7, with later cases confirmed as household-acquired by viral sequencing. Infected persons in the same household had similar nasopharyngeal viral loads (ICC=0.45, 95% CI 0.23-0.62). Households with secondary transmission had index cases with a median viral load that was 1.4 log10 higher than households without transmission (p=0.03) as well as higher living density (>3 persons occupying <6 rooms) (OR 3.3, 95% CI 1.02-10.9). Minority households were more likely to experience high living density and had a higher risk of incident infection than did white households (SAR 51% vs. 19%, p=0.01).Household crowding in the context of high-inoculum infections may amplify the spread of COVID-19, potentially contributing to disproportionate impact on communities of color

High household transmission of SARS-CoV-2 in the United States: living density, viral load, and disproportionate impact on communities of color.

BACKGROUND: Few prospective studies of SARS-CoV-2 transmission within households have been reported from the United States, where COVID-19 cases are the highest in the world and the pandemic has had disproportionate impact on communities of color. METHODS AND FINDINGS: This is a prospective observational study. Between April-October 2020, the UNC CO-HOST study enrolled 102 COVID-positive persons and 213 of their household members across the Piedmont region of North Carolina, including 45% who identified as Hispanic/Latinx or non-white. Households were enrolled a median of 6 days from onset of symptoms in the index case. Secondary cases within the household were detected either by PCR of a nasopharyngeal (NP) swab on study day 1 and weekly nasal swabs (days 7, 14, 21) thereafter, or based on seroconversion by day 28. After excluding household contacts exposed at the same time as the index case, the secondary attack rate (SAR) among susceptible household contacts was 60% (106/176, 95% CI 53%-67%). The majority of secondary cases were already infected at study enrollment (73/106), while 33 were observed during study follow-up. Despite the potential for continuous exposure and sequential transmission over time, 93% (84/90, 95% CI 86%-97%) of PCR-positive secondary cases were detected within 14 days of symptom onset in the index case, while 83% were detected within 10 days. Index cases with high NP viral load (>10^6 viral copies/ul) at enrollment were more likely to transmit virus to household contacts during the study (OR 4.9, 95% CI 1.3-18 p=0.02). Furthermore, NP viral load was correlated within families (ICC=0.44, 95% CI 0.26-0.60), meaning persons in the same household were more likely to have similar viral loads, suggesting an inoculum effect. High household living density was associated with a higher risk of secondary household transmission (OR 5.8, 95% CI 1.3-55) for households with >3 persons occupying <6 rooms (SAR=91%, 95% CI 71-98%). Index cases who self-identified as Hispanic/Latinx or non-white were more likely to experience a high living density and transmit virus to a household member, translating into an SAR in minority households of 70%, versus 52% in white households (p=0.05). CONCLUSIONS: SARS-CoV-2 transmits early and often among household members. Risk for spread and subsequent disease is elevated in high-inoculum households with limited living space. Very high infection rates due to household crowding likely contribute to the increased incidence of SARS-CoV-2 infection and morbidity observed among racial and ethnic minorities in the US. Quarantine for 14 days from symptom onset of the first case in the household is appropriate to prevent onward transmission from the household. Ultimately, primary prevention through equitable distribution of effective vaccines is of paramount importance. AUTHORS SUMMARY: Why was this study done?: Understanding the secondary attack rate and the timing of transmission of SARS-CoV-2 within households is important to determine the role of household transmission in the larger pandemic and to guide public health policies about quarantine.Prospective studies looking at the determinants of household transmission are sparse, particularly studies including substantial racial and ethnic minorities in the United States and studies with adequate follow-up to detect sequential transmission events.Identifying individuals at high risk of transmitting and acquiring SARS-CoV-2 will inform strategies for reducing transmission in the household, or reducing disease in those exposed.What did the researchers do and find?: Between April-November 2020, the UNC CO-HOST study enrolled 102 households across the Piedmont region of North Carolina, including 45% with an index case who identified as racial or ethnic minorities.Overall secondary attack rate was 60% with two-thirds of cases already infected at study enrollment.Despite the potential for sequential transmission in the household, the majority of secondary cases were detected within 10 days of symptom onset of the index case.Viral loads were correlated within families, suggesting an inoculum effect.High viral load in the index case was associated with a greater likelihood of household transmission.Spouses/partners of the COVID-positive index case and household members with obesity were at higher risk of becoming infected.High household living density contributed to an increased risk of household transmission.Racial/ethnic minorities had an increased risk of acquiring SARS-CoV-2 in their households in comparison to members of the majority (white) racial group.What do these findings mean?: Household transmission often occurs quickly after a household member is infected.High viral load increases the risk of transmission.High viral load cases cluster within households - suggesting high viral inoculum in the index case may put the whole household at risk for more severe disease.Increased household density may promote transmission within racial and ethnic minority households.Early at-home point-of-care testing, and ultimately vaccination, is necessary to effectively decrease household transmission

PRC2 disruption in cerebellar progenitors produces cerebellar hypoplasia and aberrant myoid differentiation without blocking medulloblastoma growth

Abstract We show that Polycomb Repressive Complex-2 (PRC2) components EED and EZH2 maintain neural identity in cerebellar granule neuron progenitors (CGNPs) and SHH-driven medulloblastoma, a cancer of CGNPs. Proliferating CGNPs and medulloblastoma cells inherit neural fate commitment through epigenetic mechanisms. The PRC2 is an epigenetic regulator that has been proposed as a therapeutic target in medulloblastoma. To define PRC2 function in cerebellar development and medulloblastoma, we conditionally deleted PRC2 components Eed or Ezh2 in CGNPs and analyzed medulloblastomas induced in Eed-deleted and Ezh2-deleted CGNPs by expressing SmoM2, an oncogenic allele of Smo. Eed deletion destabilized the PRC2, depleting EED and EZH2 proteins, while Ezh2 deletion did not deplete EED. Eed-deleted cerebella were hypoplastic, with reduced proliferation, increased apoptosis, and inappropriate muscle-like differentiation. Ezh2-deleted cerebella showed similar, milder phenotypes, with fewer muscle-like cells and without reduced growth. Eed-deleted and Ezh2-deleted medulloblastomas both demonstrated myoid differentiation and progressed more rapidly than PRC2-intact controls. The PRC2 thus maintains neural commitment in CGNPs and medulloblastoma, but is not required for SHH medulloblastoma progression. Our data define a role for the PRC2 in preventing inappropriate, non-neural fates during postnatal neurogenesis, and caution that targeting the PRC2 in SHH medulloblastoma may not produce durable therapeutic effects