Children with supratentorial midline pilocytic astrocytomas exhibit multiple progressions and acquisition of neurologic deficits over time

Pilocytic astrocytomas are the most common solid tumor of childhood and can arise anywhere in the central nervous system, including the posterior fossa (pf-PA), supratentorial midline (sm-PA; including optic pathway, hypothalamus, thalamus), and brainstem (bs-PA). Location (sm, bs) has been previously proposed as a prognostic factor for PA, but is difficult to separate from resection status on multivariate analysis. To overcome this limitation, we assembled a large cohort of children (n = 251) with biopsy-proved PA treated at St. Louis Children’s Hospital from 2003 – 2021 and analyzed outcomes only in patients with subtotal resection (STR; n = 81). We excluded patients with NF1, as NF1-associated gliomas often display a more indolent clinical course than their counterparts. We identified that children with STR sm-PA had a higher likelihood of multiple progressions compared to children with STR bs-PA and pf-PA. This was associated with worsening neurologic deficits over time, consistent with the sm location as a poor prognostic factor. Furthermore, the only children in our cohort with leptomeningeal dissemination or death harbored sm-PAs. Tumors in this location were also associated with an increased likelihood of non-BRAF-fusion genetic alterations and multiple oncogenic mutations. Overall, these data support location as an independent prognostic factor for PA in cases in which a gross-total resection cannot be achieved. Treating neuro-oncologists may thus wish to consider early intervention rather than watch-and-wait strategies at first progression of STR sm-PA. These patients may also benefit from earlier consideration of molecularly targeted therapy

A Systematic Screen to Discover and Analyze Apicoplast Proteins Identifies a Conserved and Essential Protein Import Factor

Parasites of the phylum Apicomplexa cause diseases that impact global health and economy. These unicellular eukaryotes possess a relict plastid, the apicoplast, which is an essential organelle and a validated drug target. However, much of its biology remains poorly understood, in particular its elaborate compartmentalization: four membranes defining four different spaces. Only a small number of organellar proteins have been identified in particular few proteins are known for non-luminal apicoplast compartments. We hypothesized that enlarging the catalogue of apicoplast proteins will contribute toward identifying new organellar functions and expand the realm of targets beyond a limited set of characterized pathways. We developed a bioinformatic screen based on mRNA abundance over the cell cycle and on phyletic distribution. We experimentally assessed 57 genes, and of 30 successful epitope tagged candidates eleven novel apicoplast proteins were identified. Of those, seven appear to target to the lumen of the organelle, and four localize to peripheral compartments. To address their function we then developed a robust system for the construction of conditional mutants via a promoter replacement strategy. We confirm the feasibility of this system by establishing conditional mutants for two selected genes – a luminal and a peripheral apicoplast protein. The latter is particularly intriguing as it encodes a hypothetical protein that is conserved in and unique to Apicomplexan parasites and other related organisms that maintain a red algal endosymbiont. Our studies suggest that this peripheral plastid protein, PPP1, is likely localized to the periplastid compartment. Conditional disruption of PPP1 demonstrated that it is essential for parasite survival. Phenotypic analysis of this mutant is consistent with a role of the PPP1 protein in apicoplast biogenesis, specifically in import of nuclear-encoded proteins into the organelle