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predominantly focuses on cardiac organoids, whereas organoid biobank could facilitate high-throughput ASO
DMD primarily manifests in skeletal muscle. Although screening across multiple disease paradigms, refining the
cardiac complications contribute to disease morbidity, identification of optimal therapeutic candidates. Further
skeletal muscle degeneration is the defining pathological advancements in next-generation ASO design could
hallmark of DMD. This underscores the necessity for improve therapeutic specificity and durability. Chemically
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developing skeletal muscle organoids and neuromuscular modified ASOs with enhanced stability, reduced off-
junction models, as well as expanding the organoid target effects, and superior cellular uptake will be critical
repertoire to encompass multi-tissue co-culture systems for sustained therapeutic efficacy. Moreover, combining
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that can enhance the translational fidelity of ASO screening. ASOs with genome-editing technologies, such as CRISPR-
To resolve this, future work should prioritize developing Cas9 could offer synergistic benefits, enabling permanent
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multi-lineage organoid systems that replicate tissue correction of pathogenic mutations alongside transient
crosstalk, such as neuromuscular junction co-cultures RNA modulation. Another crucial area of development
for muscular dystrophies. Another challenge lies in ASO is improving the organoid microenvironment to better
delivery and pharmacokinetics within organoid models. recapitulate the in vivo physiological milieu. Present
While ASOs efficiently modulate gene expression in vitro, organoid models lack immune system components, which
their therapeutic efficacy in patients depends on factors play an essential role in drug metabolism and therapeutic
such as cellular uptake, tissue penetration, and metabolic responses. The creation of immune-competent organoids
stability. The absence of a vascularized system in present incorporating patient-derived macrophages and T cells
organoid models restricts their ability to recapitulate could enable a more nuanced evaluation of ASO-induced
systemic ASO absorption, distribution, and clearance, immune activation and toxicity. In addition, the integration
limiting their predictive accuracy for clinical applications. of artificial intelligence (AI-driven high-throughput
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The development of vascularized, perfusable organoids screening methodologies could accelerate ASO candidate
or microfluidic-based organ-on-a-chip models represents optimization, enabling predictive modeling of patient-
a promising avenue to overcome these limitations, specific therapeutic outcomes and refining personalized
enabling a more physiologically relevant assessment treatment selection.
of ASO pharmacodynamics. In addition, genetic and
epigenetic variability among PDOs poses a challenge The convergence of PDO technology with ASO
for reproducibility. iPSC-derived organoids may exhibit screening marks a paradigm shift in precision medicine,
batch-to-batch variability, which can affect experimental offering scalable, physiologically relevant, and patient-
specific disease models for therapeutic evaluation. The
consistency and ASO responsiveness. Ensuring genetic study by Means et al. provides compelling evidence for the
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stability through standardized differentiation protocols,
whole-genome sequencing, and epigenetic profiling is feasibility of PDO-based ASO screening as an accelerated
paramount to improving reproducibility and ensuring and effective platform for gene therapy validation. However,
therapeutic reliability. The final barrier to widespread challenges remain, particularly in tissue representation,
clinical adoption is regulatory approval and integration ASO pharmacokinetics, and clinical integration.
into existing drug development pipelines. While the Addressing these limitations through the expansion of
study establishes a robust pre-clinical platform, regulatory organoid models, optimization of ASO therapeutics, and
agencies have yet to fully incorporate organoid-based incorporation of advanced bioengineering and AI-driven
drug screening into standardized approval frameworks. technologies will be pivotal in establishing PDOs as the
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Furthermore, ethical considerations surrounding patient- gold standard for personalized medicine. With continued
derived iPSC biobanking, data privacy, and long-term advancements, organoid-based precision medicine is
storage of PDOs require careful oversight. Developing poised to redefine drug discovery, revolutionize gene
comprehensive regulatory guidelines and quality control therapy development, and transform clinical treatment
measures will be instrumental in facilitating the transition paradigms for genetic disorders.
of PDO-based drug screening into routine clinical use. Acknowledgments
To fully harness the therapeutic potential of PDO-
based ASO screening, future research should focus None.
on broadening organoid models, optimizing ASO Funding
therapeutics, and enhancing clinical translation strategies.
One particularly promising direction is the extension None.
of this methodology to other genetic disorders, such as
spinal muscular atrophy, Huntington’s disease, and cystic Conflict of interest
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fibrosis. The establishment of a diverse, patient-specific The authors declare no conflicts of interest.

Volume 1 Issue 3 (2025) 3 doi: 10.36922/OR025120012