Test Reaserches roadmap

Segment 1 - DRUG REPURPOSING

Parallelized, AI-informed Drug Repurposing

Our search for 8p treatments anchors in high-throughput, bias-minimized phenotype-rescue drug repurposing, in three 8p-hero-derived tissue types: fibroblasts (for baseline structural/functional and cross-tissue insights); iPSC-differentiated neural progenitor cells (for lineage-specific developmental insights); and iPSC-differentiated excitatory neurons (for insights on differentiated tissue dysfunction). In parallel, we aim to leverage ancillary insight from patient-reported outcomes, partnered academic projects (some using machine learning to mine multimodal findings), and data-grounded, physician-initiated off-label n-of-1-or-few trials.

Starting with pilot findings from an inv/dup/del family trio, 8p-hero-derived primary fibroblasts will be assayed with a standardized, validated panel of dyes targeting mitochondria, nuclei, and other organelles/cell compartments (cell painting). Growth under challenge (e.g., on galactose without glucose, or in presence of DNA damaging agents) will help identify potential rescue drugs. Ideally, many assessed phenotypes in 8p fibroblasts may statistically differ in patients versus controls – perhaps with one or more robust mitochondrial and/or nuclear disease phenotype that can be ameliorated in a 96- or 384-well-plate screening assay optimized in the lab of Professor Nicoleta Moisoi at the Leicester School of Pharmacy. The Broad Repurposing Hub (REPO) libraries and/or ReFRAME collection could then be screened Read more…

Segment 2 - dISEASE MODELING

Disease in a Dish & Therapeutic Target Assessment

The advantage of unbiased drug repurposing screens is that they don’t require complete knowledge of the underlying disease mechanism. As we’ll discuss in Segment 3, targeting root causes of 8p disorders entails fixing and/or replacing damaged chromosomes, which drive chronic tissue stress even after neurodevelopment. The goals of this Segment are discovery of disease-modifying drug targets and aneuploidy-compensating pathways like the integrated stress response (ISR), which will provide added and in some cases significant therapeutic benefit above what will be provided by repurposed drugs.

We envision disease modeling using three 8p-hero-derived cell types of increasing complexity: iPSC-differentiated neural progenitor cells (NPCs), iPSC-differentiated excitatory neurons (iNeurons), and iPSC-differentiated brain organoids. Read more…

Segment 3 - Cell and Chromosome Therapies

Building the foundational toolkit

As both a hedge and a complement to Segments 1 and 2, we recommend collaborating with multiple academic groups (and if possible early-stage biotech startups) on current and emerging modalities that are CNV-agnostic. As stated several times above, the rationale for focusing on the root cause of 8p disease – damaged 8p chromosomes – is that it obviates the need to validate driver genes one by one.

As an initial down payment on the promise of chromosome therapy, Project 8p will sponsor a diversified five-pronged approach in the lab of Professor Jason Sheltzer at Yale University in order to identify techniques that restore 8p disomy (i.e., a normal diploid karyotype) in the pathfinding inv/dup/del pioneer proband. The first approach involves simply growing 8p iPSCs in the lab over many generations (cell divisions) and checking for spontaneous loss of the damaged 8p chromosome, as has been previously demonstrated to happen with Down Syndrome iPSCs. The second approach is an accelerated form of the first approach that involves treating 8p iPSCs with a drug that increases the rate of spontaneous loss of damaged 8p chromosomes. The third approach uses CRISPR (gene editing) to target DNA sequences specific to damaged 8p chromosomes, resulting in their physical elimination from the cell. The fourth approach also takes advantage of CRISPR but instead of targeting naturally occurring sites unique to damaged 8p chromosomes, a susceptibility gene is inserted into the damaged 8p chromosomes of 8p iPSCs, rendering these edited cells sensitive to a drug that selectively kills them. The fifth approach involves a technique called microcell-mediated chromosome transfer (MMCT) whereby a second copy of a normal 8p chromosome is delivered to 8p iPSCs, followed by application of the four preceding approaches on trisomic post-MMCT 8p iPSCs to restore a normal karyotype. Read more…

Segment 4 - In hero Studies

In Hero Studies To Elucidate Disease Drivers And Prototype Augmentative Terapies

Project 8p will continue to fund efforts to identify and recruit 8p heroes into the IRB-approved Chromosome 8p Registry and Biorepository led by principal investigator, Bina Shah, CEO of Project 8p in an effort to be agile with a sense of urgency. This is the start of a robust and standardized retrospective and prospective natural history study.

Project 8p will engage with cutting-edge biotech startups developing brain-computer interface devices, neuromodulation, or assistive technologies to explore pilot programs that will give 8p heroes access to experimental augmentation therapies. Read more…

Team Scientists

Dr. Alysson Muotri

A Brain Organoid model op 8p neurodevelopment

A Brain Organoid model op 8p neurodevelopment
A model for probing genes, pathways, networks and screening therapeutics.
Differentiation of 8p family trio iPSCs into cortical brain organoids.

Dr.Aryeh Warmflash

Germ layer patterning defects in
8p.

Understand the impact of 8p genes and regions on changes in early embyo developmetal.
Identify potential points of early therapeutic intervention
Assess germ layer patterning and changes in gene expression during early development in 8p lines

Dr. Hiruy Meharena

Omics characterization of neurodevelopmental defects in 8p

Understand how 8p CNVs affect different brain cell types.
Identify phenotypes for high throughput screening.
Identify therapeutic intervention points.

Dr.Stefan Printer

CRISPR activation and inhibition lines for investigation of 8p candidate genes.

Tools for probing candidate gene function in a dose- controlled manner.
Tools for top-down and bottom-up screening of 8p modifiers
Create euploid line with inducible CRISPRa/ and NGN2

Dr.Jason Sheltzer

Chromosome engineering for restoring disomy in 8p

Understand the viability of chromosome engineering as a therapeutic strategy for 8p.
Create genetically matched 8p and control lines.
Test five unique strategies for restoring chromosome 8 disomy in 8p iPSCs.