EyePoint’s quest to reduce treatment burden for patients with age-related macular degeneration has hit a bump in the road, with a clinical miss driving the biotech’s stock down by 70%.

EyePoint’s late-stage eye candidate has failed to beat approved treatment aflibercept in improving vision for patients with wet age-related macular degeneration, putting pressure on an upcoming readout for the same asset.

Lugano, a Phase 3 trial of about 400 patients, tested EyePoint’s investigational tyrosine kinase inhibitor (TKI) duravyu against the standard-of-care aflibercept, a vascular endothelial growth factor (VEGF) inhibitor marketed by Regeneron Pharmaceuticals as Eylea and also sold by a handful of biosimilars makers. The trial’s main goal was improving average change in vision after two years, as measured by best corrected visual acuity (BCVA) using an eye chart.

The primary endpoint wasn’t met and EyePoint failed to share any other information around the miss in a Monday press release. Subsequently, the biotech’s stock crashed 70% at market open, falling to $4.28 per share as of 10 a.m. ET.

In a presentation for an accompanying investor call, EyePoint revealed that the BCVA mean change from baseline was above five points for aflibercept, as compared to a change between zero and five points for duravyu. Again, the biotech didn’t share specific figures behind the data.

Instead, the company touted findings from an ad hoc analysis that removed nine of 211 patients receiving the investigational treatment who had experienced vision loss of 15 or more letters “for reasons unrelated to wAMD.”

The company said these patients “confounded” a primary endpoint win, adding that when they were removed, duravyu was non-inferior to aflibercept. None of the patients in the aflibercept arm experienced vision loss equal to or greater than 15 letters that was unrelated to AMD.

The Massachusetts-based biotech noted that 3–5% of patients taking aflibercept typically have lost 15 or more BCVA letters in past Phase 3 trials, implying an unusually clean—and possibly over-performing—control arm, according to Mizuho analysts. That framing is “plausible,” the analysts said in a Monday note, but questions remain about what caused vision loss for the nine patients receiving duravyu.

EyePoint also trumpeted “positive” secondary endpoints, such as a 42% reduction in treatment burden, as compared to Mizuho analysts’ 30–35% expectation. The finding was superior to Eylea, equating to almost two fewer injections through week 56, according to EyePoint.

That improvement contributes to “excellent” secondary-endpoint data, Mizuho wrote Monday, while calling the overall readout “mixed.” Other secondary data include a clean safety profile and high supplement-free rates for patients on duravyu up to week 32.

Additional details from Lugano are expected to be shared at the Retina Society 59th Annual Scientific Meeting in Los Angeles in late September.

Now, all eyes turn to Lugano’s twin trial, Lucia. A clean win in the identically designed late-stage study would “substantially de-risk the ad hoc narrative” and support EyePoint’s new drug application filing that is planned for the first half of next year, Mizuho wrote.

“A second miss would be far more problematic,” the analysts wrote.

“The secondaries reinforce duravyu’s differentiated durability and safety,” Mizuho explained. “But the primary miss is a genuine overhang we can’t fully dismiss until LUCIA.”

Topline data from Lucia are expected in October.

EyePoint’s mixed readout is good news for Ocular Therapeutix, a direct competitor touting its own TKI candidate called axpaxli for wet AMD. Both biotechs aim to reduce the frequency, and therefore burden, of anti-VEGF injections.

After reporting Phase 3 findings earlier this year, Ocular said recently that a new drug application was on track to be submitted to the FDA in the fourth quarter of this year. The late-stage trial met its primary endpoint, with 74.1% of subjects in the axpaxli arm maintaining vision at week 36, a statistically significant 17.5% risk difference compared to aflibercept.

Ocular opened up more than 6% on Monday at $10.72 per share.

Johnson & Johnson secured rights to prizlon-cel in 2023 for $245 million upfront. The asset in June last year showed a 100% response rate as a second-line option in a Phase 1b study for 10 patients with relapsed or refractory large B cell lymphoma.

Johnson & Johnson has given up on an investigational CAR T therapy for large B cell lymphoma that the pharma had previously licensed from AbelZeta Pharma.

Rights to the asset, dubbed prizloncabtagene autoleucel (prizlon-cel), were returned to AbelZeta in July, the U.S.- and China-based company said in a Sunday release. Prizlon-cel is being developed for the treatment of third- or later-line large B cell lymphoma (LBCL).

J&J through its former Janssen subsidiary fronted $245 million in May 2023 to secure the exclusive right to develop and commercialize prizlon-cel—alongside other CAR T assets—outside the Greater China region. The pharma at the time also agreed to pay certain development, regulatory and sales milestones, though these remain undisclosed.

A few months later, in December that year, the companies amended the agreement to give J&J the option to obtain the exclusive rights to prizlon-cel in China. The financial terms of this arrangement were likewise kept under wraps.

BioSpace has reached out to J&J for more details regarding the decision to return prizlon-cel to AbelZeta and the status of the partnership. The asset no longer appears on J&J’s pipeline page.The pharma listed prizlon-cel—which it labeled JNJ-4496—in its full-year 2025 report in January, but the molecule no longer appeared in the company’s first quarter presentation in April.

In June last year, J&J released Phase 1b data showing that prizlon-cel elicited a treatment response in all dosed patients with relapsed or refractory LBCL who had undergone one prior line of therapy. Eight of the 10 patients showed a complete response. For the 12 patients receiving the candidate as a third-line treatment, overall response rate was 92% and complete response rate was 75%.

Analysts from Truist Securities at the time called these outcomes “compelling,” representing what they said would be an “encouraging step up” from Gilead’s Yescarta, currently the standard of care in LBCL. Yescarta’s complete response rate is 65% in the second-line setting, according to its label.

The decision to divest prizlon-cel comes as J&J buffs up other areas of its CAR T portfolio. Late last month, the pharma paid $785 million upfront—and earmarked up to $140 million—to partner with Sail Biomedicines to advance in vivo CAR T therapies for immune indications. The agreement also gives J&J the option to outright acquire Sail in the future for $2.58 billion.

Despite ranking among the world’s top scientific producers, Italy has struggled to turn research into startups. Now, stronger tech transfer offices, private capital and returning scientists are pushing the country toward a more mature innovation economy.

Italy is finally beginning to convert its highly regarded scientific advantages into real companies.

After years of world‑class research output, superb clinical excellence and a growing venture capital (VC) base, there is optimism for Italian startups becoming increasingly global players, said several Italian VCs that spoke to BioSpace.

Tech transfer offices (TTOs) have improved. Italians are moving back from abroad, nurturing the talent ecosystem. Although cultural resistance to commercialization remains strong among scientists, consistency in funding sources such as incubators and VC backing bodes well for the future of Italian innovation.

Italian research and clinical firepower

Italy has long had high-standing research. According to the Nature Index, which tracks high‑quality scientific publications across 178 top journals, Italy ranks tenth globally and fourth in Europe for overall research output.

In terms of hubs, the Lombardy region, with Milan as its capital, has historically driven innovation as the financial center of the country, said Fabrizio Calisti, medical director at the Rome-based Angelini Ventures. Milan also has a strong science research base, he said, but excellent universities exist across the country.

In Italy, many startups come out of university research, added Lorenzo Pradella, board director at Milan-headquartered Zcube. To that benefit, TTOs have become more efficient, said Calisti and Diana Saraceni, founder and managing partner at Milan-based Panakès Partners.

As discussed in previous Insights articles and Denatured podcasts on the European financing system, expedient TTOs are crucial for company creation, attracting foreign capital, retaining value inside a country and scaling companies without losing a national identity.

Unfortunately, both Saraceni and Calisti agreed that the mindset of startup creation out of science research is still not very popular. “Researchers tend to keep researching for the sake of it,” Saraceni said, adding that scientists often feel it’s unethical to make money.

Investment encouragement

Despite the researcher mindset, Saraceni expressed optimism about increased investment volumes coming into Italian and European companies each year. The trend builds on a strong foundation started by the Cassa Depositi e Prestiti (CDP), Italy’s national promotional institution and public development bank, about 10-15 years ago, she explained. CDP supported creation of incubators as well as backed several VCs, underscoring a financial and science ecosystem that attracts international capital as well.

Although CDP catalyzed the ecosystem, capital today is overwhelmingly private, Saraceni said. Italy now has about a half-dozen local VCs active in biotech, she said, with several VCs having a corporate VC arm. The VC ecosystem is growing and connected better with other European players, Calisti added.

On the national level, Italy’s multibillion National Recovery and Resilience Plan, funded through the E.U., also has offered a fraction of the funds to boost biotech, medtech and TTO investment, Calisti said.

Italy’s ecosystem is also aided by “brain repatriation:” more natives are returning home, bringing talent and company expertise they gained outside the country, Calisti and Saraceni said. Tax incentives have also drawn non-Italians in the sciences to move, Saraceni added.

A key challenge for Italian companies will be to scale while maintaining the Italian footprint, Calisti said. Relocation to bigger hubs can be normal, but it’s important for the Italian ecosystem to retain value, he added. Manufacturing also has a strong based in Italy, Calisti said, which could hold an important element to keep some Italian roots.

Still, Italian companies should consider thinking themselves as European-first, Saraceni said.

The cultural resistance to commercialization won’t disappear overnight, but investors argue the momentum is real: more founders, more capital and more global connectivity. With manufacturing strength and a growing European orientation, Italy may be on the edge of transforming its scientific reputation into a consistent pipeline of globally competitive biotech companies.

Taiho Pharmaceutical and Cullinan Therapeutics’ cancer asset is already under FDA review in a later setting, with a decision expected by February.

Taiho Pharmaceutical and Cullinan Therapeutics’ candidate zipalertinib significantly improved progression-free survival for patients with advanced non-small cell lung cancer in a late-stage trial. Already under FDA review for previously treated NSCLC, the new results tee up a potential FDA filing as a first-line treatment.

The partners said in a Wednesday night release that REZILIENT3, a global Phase 3 study for patients with NSCLC harboring epidermal growth factor receptor (EGFR) exon 20 (ex20) insertion mutations, met its primary endpoint at an interim analysis.

The companies did not share figures behind the PFS win but said its experimental EGFR inhibitor plus chemotherapy demonstrated a statistically significant and clinically meaningful improvement in PFS when compared to chemotherapy alone.

The risk of disease progression or death is measured using a hazard ratio, with any ratio under one demonstrating that the investigational treatment is tied to a better PFS than those in the comparative arm. The closer the hazard ratio is to zero, the better the survival rate is.

An independent data committee recommended Taiho and Cullinan unblind REZILIENT3, suggesting “the magnitude of results could be significant,” according to Leerink Partners. “The total trial was powered to detect a hazard ratio of 0.60 at 90% power,” William Blair analysts wrote in a Thursday note, “strongly suggesting the interim analysis came in below 0.6 to achieve stopping criteria.”

A hazard ratio potentially below 0.6 means the zipalertinib-chemo combo could compare favorably to Dizal Pharmaceutical and AstraZeneca’s EGFR inhibitor Zegfrovy, which demonstrated a hazard ratio of 0.65—or a 35% improvement in PFS—versus chemotherapy in a Phase 3 first-line NSCLC study.

The full data will be shared at an upcoming medical conference, and the partners plan to seek regulatory approval in the first-line setting, they said in their announcement.

Zipalertinib is currently under FDA review as a monotherapy in previously treated EGFR ex20 NSCLC, with a decision expected by Feb. 27.

William Blair assumes the REZILIENT3 findings will be filed as a supplemental new drug application immediately if the approval for previously treated patients goes through, adding that the partners will likely make regulators aware of the new data to see if it can be included in the current review.

While patients with EGFR ex20 NSCLC account for a relatively small portion of the market, William Blair believes “zipa’s strong efficacy and tolerability could prove beneficial in the first-line setting.” The safety profile for the investigational arm in REZILIENT3 was manageable, the partners said.

If approved, Taiho-Cullinan’s zipalertinib-chemo combo would be the third first-line therapy to hit the market in the indication, joining Zegfrovy and Johnson & Johnson’s bispecific antibody Rybrevant plus chemotherapy, the latter of which carries certain tolerability issues, William Blair noted.

AstraZeneca recently scooped up commercial rights to Zegfrovy for up to $1.5 billion, suggesting that the drug will soon be commercialized in the U.S., the analyst said. The drug secured an accelerated approval from the FDA last year, but China-based Dizal hasn’t been able to get commercialization efforts off the ground since then.

Rybrevant and chemotherapy demonstrated a median PFS of 11.4 months in the first-line setting, while Zegfrovy showed a 10.3-month median PFS. Therefore, Leerink analysts think zipalertinib will need to show a median PFS around 11 months or more to compete. Anything below 11 months may not justify the added chemotherapy toxicity compared to Zegfrovy monotherapy, Leerink wrote.

Taiho and Cullinan came together in 2022, with the former acquiring a Cullinan subsidiary housing zipalertinib for $275 million upfront.

Cullinan now has the chance to make up to $130 million in regulatory milestones, plus a 50/50 profit share in the U.S.

The terms of the agreement with Royalty Pharma suggest the market potential of Zealand Pharma’s polycythemia vera drug candidate may be undervalued, according to Jefferies analysts.

In a royalty purchase and sale pact with Royalty Pharma, Zealand Pharma has exchanged most of its economic interest in the investigational polycythemia vera drug rusfertide that it had once developed with Protagonist Therapeutics.

Under the terms of the agreement, announced Wednesday night, Zealand will get $50 million upfront and stands to receive up to $50 million more on the first anniversary of the deal’s closing. In exchange, Zealand will surrender most of its remaining economic interests in rusfertide—those it hadn’t already returned to ex-partner Protagonist—including regulatory and sales milestones, as well as certain royalties on yearly product sales.

In 2012, Zealand partnered with Protagonist to advance disulfide-rich peptide therapies, one of which would eventually be rusfertide. The deal leveraged Protagonist’s platform for targets of Zealand’s choosing, Genetic Engineering & Biotechnology News reported at the time. Zealand was in charge of all preclinical and clinical work, though Protagonist retained the option to co-finance and co-develop the resulting assets.

The collaboration was terminated in 2014, kicking off a years-long dispute about payments still owed to Zealand. The two companies came to a resolution in August 2021, with both parties agreeing to a reduced milestone structure, plus royalties, though specifics of this new arrangement remain undisclosed.

In 2024, Protagonist partnered with Takeda to develop and commercialize rusfertide. Takeda and Protagonist in March announced that the FDA accepted their new drug application for rusfertide, granting it priority review. A decision is expected in the third quarter of this year.

Under the Wednesday deal with Royalty, Zealand will only retain the right to 0.25% royalties on global sales of rusfertide in excess of $1.5 billion—a stipulation that Jefferies said shows that Zealand expects rusfertide’s future sales to exceed current forecasts.

The terms of the Royalty deal suggest “consensus may underestimate the drug’s commercial potential,” the firm told investors late on Wednesday, pointing to the approximately $1.2 billion forecast by Takeda’s 2034 fiscal year.

“While the undisclosed milestones prevent a precise valuation analysis, Zealand’s decision to retain royalty exposure only above $1.5bn suggests it sees a realistic path to sales exceeding that level,” Jefferies said. The group has high expectations for rusfertide, pegging peak sales to hit around $2 billion.

Designed to be given subcutaneously every week, rusfertide works by mimicking the hormone hepcidin, which helps regulate the production of red blood cells. This mechanism addresses the underlying iron dysregulation in polycythemia vera, in turn lowering the excess production of red blood cells that is characteristic of the blood cancer, according to Zealand’s Wednesday release.

Phase 3 data released in March last year showed a 77% response rate in patients on rusfertide versus 33% in placebo comparators. Participants were on background treatment with standard of care. Rusfertide likewise significantly boosted the proportion of patients who didn’t need phlebotomy from weeks 20 through 32.

In biomanufacturing, scale has long been synonymous with success. Bigger bioreactors, larger facilities, and expanded footprints traditionally defined the path to higher output. But that paradigm is shifting with intensified bioprocessing. Today, the industry is embracing a more nuanced, efficient approach—one that prioritizes productivity over size, agility over rigidity, and integration over segmentation. Intensified bioprocessing is not just an incremental improvement; it is a fundamental rethinking of how biologics are made.

“Intensified bioprocessing aims to improve the productivity and efficiency of biomanufacturing,” explains Julie Kozaili, PhD, principal scientist at Asahi Kasei Bioprocess. “This is often achieved by designing new processes or modifying existing ones to increase output per unit time or equipment volume.”

That deceptively simple definition captures a sweeping transformation. Instead of relying on traditional batch processes, intensification often involves running at higher cell densities, integrating multiple process steps, and transitioning toward continuous or semi-continuous operations.

The implications are significant. Intensified processes can reduce facility size, minimize resource consumption, and shorten development timelines—all while maintaining or even improving product quality. For an industry under constant pressure to deliver therapies faster, these advantages are hard to ignore.

The urgency behind intensification is driven by both scientific and economic realities. Many modern therapeutics—particularly viral vectors and gene therapies—face inherent production challenges. Low yields, complex manufacturing requirements, and stringent quality standards make scaling difficult and expensive.

In viral-vector development, one of the central bottlenecks is simply producing enough material. Clinical applications often require a minimum effective dose volume, yet production systems struggle to generate sufficient yield, forcing manufacturers to concentrate limited output into small delivery formats. Legacy adherent cell culture technologies compound the problem by relying on scale-out strategies—adding more units rather than increasing efficiency—making cost reductions difficult as production expands.

Intensified bioprocessing offers a different path. It “is important because it allows manufacturers to increase capacity without new facilities, reduce equipment footprint, reduce media, buffer, and utility usage per gram of product, and shorten scale-up, tech transfer, and time-to-clinic timelines,” Kozaili says.

For companies working with unstable or complex molecules, speed can be just as important as scale. Faster processing reduces the risk of degradation and accelerates the path from development to commercialization.

Beyond cost: speed and flexibility

Although cost savings are often cited as a benefit of intensification, industry leaders emphasize that its true value lies beyond the cost of goods. “Intensified bioprocessing is less about driving down cost and more about enabling speed, flexibility, and fit,” says Mark Schofield, PhD, director of science at Cytiva. “For monoclonal antibodies in particular, the industry’s priorities are getting to launch faster, making better use of existing facilities, and being able to respond to uncertain or fluctuating demand.”

This shift in perspective reflects broader changes in the biopharmaceutical landscape. Pipelines are increasingly diverse, with smaller patient populations and more specialized therapies. Manufacturing systems must be adaptable, capable of switching between products or scaling production up and down as needed. “Intensification helps companies do all three by rethinking how processes are designed and scaled,” Schofield adds.

Companies such as Repligen are advancing upstream intensification through perfusion-based systems designed to sustain high cell densities and continuous productivity. Perfusion cell culture, a cornerstone of many intensified strategies, continuously feeds fresh media while removing waste and product, allowing cells to remain in an optimal growth state over extended periods. This approach not only improves yield but also creates a more stable and controlled production environment compared to traditional fed-batch methods. Repligen’s filtration and analytical technologies further support this shift by enabling continuous clarification and real-time monitoring, helping bridge the gap between process development and scalable manufacturing.

Beyond large platform providers, a growing number of specialized innovators are helping push intensified bioprocessing forward, particularly in high-demand areas like viral-vector manufacturing and upstream control.

Meanwhile, Batavia Biosciences is tackling one of the most persistent challenges in gene therapy: low viral-vector yields. Traditional adherent cell culture systems often require scaling out—adding more equipment rather than increasing efficiency—which drives up costs without significantly improving productivity. Batavia’s intensified approach centers on integrated solutions that combine optimized cell lines, streamlined purification processes, and novel bioreactor designs to dramatically increase output. By enabling higher yields within a smaller footprint, these strategies effectively miniaturize manufacturing, making it possible to produce clinical and commercial quantities without the need for large-scale facilities.

Together, these efforts underscore a key theme in intensified bioprocessing: innovation is not confined to a single step or technology. Instead, it is emerging across the entire workflow, from upstream cell culture to downstream purification and process analytics.

Real-world applications

The promise of intensified bioprocessing is being realized through a growing ecosystem of technologies. Asahi Kasei Bioprocess, for example, has developed solutions that support intensification at multiple stages. “We support intensified bioprocessing across upstream and downstream operations,” Kozaili explains, pointing to innovations such as hollow-fiber microfilters for high-intensity cell culture clarification and advanced virus filtration systems designed for continuous processing.

These technologies are engineered to handle the increased throughput associated with intensified upstream processes. High-density cultures generate larger volumes of product, which must be efficiently clarified, purified, and stabilized without compromising quality.

Downstream, continuous virus filtration systems can operate at low flux over extended periods while maintaining robust viral clearance. Inline buffer formulation systems further streamline workflows by eliminating the need for large storage tanks and ensuring consistent buffer quality in real time.

Automation and integration are also key components. New ultrafiltration and diafiltration systems are being designed for flexibility, allowing them to be deployed upstream or downstream and enabling seamless process integration.

Designing for intensification

Though technology is a crucial enabler, successful intensification requires more than just new equipment. It demands a holistic approach to process and facility design. “At CRB, our role is to help clients translate emerging process concepts into facilities that are safe, operable, and scalable,” says John Rubero, senior fellow in purification bioprocessing.

One of the defining characteristics of today’s intensification efforts is that they are often partial or hybrid implementations. Fully continuous, end-to-end processes remain relatively rare. Instead, manufacturers are adopting elements of intensification—such as integrating continuous perfusion with multi-column capture chromatography—within otherwise traditional workflows. This incremental approach allows companies to realize benefits without fully overhauling their operations. It also provides a pathway for future evolution as technologies mature.

Despite its advantages, intensified bioprocessing is not without challenges. One of the most significant is bridging the gap between process development and commercial-scale implementation. “While the practice of linking unit operations together is largely accepted, real-time control of an end-to-end continuous process remains challenging,” Rubero explains.

In traditional batch processes, control strategies are relatively straightforward because lot traceability is easy to maintain. But intensified systems—especially continuous ones—require real-time monitoring and advanced control strategies to ensure process stability and product quality.

“It is not realistic or necessary to find and assign a sensor to monitor each critical process parameter or critical quality attribute,” Rubero says. “Instead, a combination of direct measurements, soft sensors, multivariate models, and process understanding is required for effective process control.”

So, the industry is moving toward integrated approaches that combine process analytical technology (PAT) with mechanistic and data-driven models. These systems enable more sophisticated monitoring and control but are still evolving in terms of reliability and adoption.

Operational barriers

Technical challenges are only part of the equation. Intensification also requires a shift in mindset—one that can be difficult for organizations accustomed to established manufacturing paradigms. “In many cases, the technologies are either new or have novel applications, creating a learning curve,” Kozaili acknowledges.

Training gaps, operational changes, and resistance to new approaches can slow adoption. Teams must adjust not only their processes but also their thinking, moving away from long-standing practices toward more dynamic, integrated systems.

As Schofield notes, “adopting new approaches inevitably comes with skepticism.” Externally, there can be hesitation to move away from established technologies. Internally, organizations might question how intensified solutions might impact existing product lines. Those discussions, however, are part of the transition.

Despite these challenges, momentum is building. As intensified technologies demonstrate their value in real-world applications, resistance is gradually diminishing. “Over time, evidence and adoption speak for themselves,” Schofield says.

Kozaili emphasizes the importance of organizational alignment. “We had to change the company’s established mindset by securing support to develop these technologies and clearly show the value of these approaches,” she explains.

Collaboration also plays a key role. For technology providers, working closely with customers to test and refine solutions helps build confidence and accelerate adoption. “For our customers, it’s about finding the right partners to test the technologies, while providing appropriate feedback for improvement,” Kozaili adds.

Looking ahead, the trajectory of intensified bioprocessing is clear. Purpose-built facilities designed specifically for intensified operations will become more common, replacing retrofitted batch plants that struggle to accommodate new workflows, because intensified bioprocessing is no longer a niche concept reserved for early adopters. It is rapidly becoming a central pillar of modern biomanufacturing strategy.

A multi-disciplinary, multi-institutional group of researchers said they relied on their expertise in genetics, mechanics, chemistry, and biology to create a chip the size of a postage stamp to model a particular class of heart conditions.

The team, led by Guang Li, PhD, an associate professor in the University of Pittsburgh School of Medicine’s department of cell biology, has grown heart valves on organoids. The study “Human iPSC-derived heart valve-like assembloids model valve development and disease pathology” appears in Cell Stem Cell and is an important step toward better understanding and treating a number of serious heart disorders, according to the scientists.

This kind of research often depends on animal models, where researchers can study the development of heart valves that grow much quicker than those of humans (which take nearly 10 weeks to fully develop), and don’t raise the same ethical dilemmas as it would in humans. But, Li said, “human valves are very different from animal valves.” Imagine the physiological and genetic differences between a person and, for instance, a zebrafish. “To study human valve diseases, we need human valve models.”

The organoids were grown from pluripotent, adult human stem cells, which can be generated from skin, blood, or other cells, then coaxed into developing into cells from a body part of interest; in this case, a human heart. Different types of organoids can be combined into “assembloids” to better model complex organs that natively originate from combinations of different tissues.

However, a functioning heart is more than a cluster of certain types of cells. Its development and continued operation are dependent, among other things, on a complex interaction of different forces. To build analogs of those forces into the model, Li sought the engineering expertise of colleagues, including Lance Davidson, PhD, the William Kepler Whiteford Professor of bioengineering in the Swanson School of Engineering and Si-Yang Zhen, PhD, a professor of biomedical engineering at Carnegie Mellon University.

“This kind of project is really a hallmark of the community of researchers in Pittsburgh,” Davidson said.

Valve grown on heart assembloid surface
To create a model, Li grew a valve on the surface of a heart assembloid. Then the team stimulated growth by designing ways to mimic the forces that would act on an embodied heart, a flowing medium to simulate blood, an endothelial culture which simulates cells that line heart valves, and even a set of magnetized beads that moved according to the placement of a magnetic belt to simulate muscle contraction.

With the organoid working to simulate a heart with valves, the team now had a model they could use to study four types of valve disorders, including mitral valve prolapse (MVP), a genetic disorder affecting seven to eight million individuals in the US at any given time.

When Li introduced a mutation associated with the disease, the developing valves showed signs of MVP. In other cases, damage was simulated or introduced to mirror the damage that can occur to a person’s valves throughout life in conditions such as valve calcification; cryo-injury; and complications from hypoglycemia and diabetes.

Li was able to begin studying the organoids, identifying some pathways responsible for the development problems associated with MVP and ways they can be corrected. He was also able to develop models for the acquired deficiencies and will go on to look for ways to treat them.

Next, however, Li plans to add complexity to his assembloids, growing them with two chambers and growing the valves inside them, instead of on the surface, to better model a real human heart.

A team of researchers at Vanderbilt Health and the Translational Genomics Research Institute (TGen) has used image-based spatial transcriptomics to profile cellular programs involved in heart transplant rejection, an analysis the group says could help sharpen diagnosis, predict treatment response, and stratify long-term risk after transplantation.

The study, “Dynamic cellular programs of human cardiac allograft rejection revealed by spatial transcriptomics,” was published recently in Nature Cardiovascular Research. In it, the investigators applied image-based spatial transcriptomics to longitudinal human endomyocardial biopsy samples from 62 adult and pediatric heart transplant recipients during and after histologically diagnosed rejection.

Allograft rejection remains a major challenge after solid organ transplantation, with up to approximately 40% of recipients experiencing rejection within one year after transplant, and can contribute to long-term graft failure and death. In heart transplantation, clinicians routinely monitor for rejection using endomyocardial biopsies, with current diagnostic approaches relying heavily on histology. However, histologic findings do not always reflect a patient’s clinical course: “clinical presentation varies dramatically within the same grade of histologic rejection” and “response to antirejection therapy is heterogeneous, including lack of response in some patients,” the authors wrote.

“We see substantial variability in histologic rejection grades, which impacts the precision of our immunosuppressive therapies, with a potential for over- or under-immunosuppression with downstream clinical consequences,” said co-senior author Ravi Shah, MD, the Gottlieb C. Friesinger II professor of cardiovascular medicine and professor of medicine at Vanderbilt Health. “We directly examined molecular phenotypes in tissue during rejection and antirejection therapy after heart transplantation to start to understand this heterogeneity.”

The researchers analyzed longitudinal biopsy samples collected from the same patients during acute rejection and after various immunomodulatory therapies. Using spatial transcriptomics, they mapped gene expression across tissue architecture at subcellular resolution and identified 28 cell types, including immune and parenchymal cells, that differed across rejection classes. The team found broad overlap in transcriptional states across rejection severity, as well as substantial molecular heterogeneity within the same rejection grades—variation that was not apparent by histology alone.

Baseline rejection biopsies also differed between patients who responded to augmented immunomodulatory therapy and those who did not. In the paper, the authors reported that nonresponders showed “baseline T cell hyperactivation and tissue remodeling genes,” suggesting that molecular profiling could eventually help distinguish patients likely to benefit from standard approaches from those who may need more intensive or alternative therapies.

The analysis also linked cell-specific gene expression patterns to cardiac allograft vasculopathy (CAV), a chronic form of rejection that limits long-term survival after heart transplantation. “The molecular heterogeneity may help explain the spectrum of clinical presentations—from complete lack of symptoms to cardiogenic shock—for the same histologic grade of rejection,” said co-corresponding senior author Nicholas Banovich, PhD, vice president of scientific development and professor at TGen. “We expect that data generated through approaches like ours will inform early biomarker and drug discovery to meaningfully prolong transplanted organ survival.”

Together, the findings suggest that spatial transcriptomics could add clinically relevant molecular context to conventional biopsy interpretation, helping researchers better subtype rejection, predict therapeutic response, and identify patients at risk for long-term complications such as CAV.

Since its discovery in 2009, Candida auris—a multidrug-resistant pathogenic yeast—has caused deadly outbreaks around the world and is responsible for roughly 3,000 deaths in patients in hospitals and long-term care facilities per year in the U.S. The fungus is known to colonize human skin, however, the mechanisms that it uses to persist on skin remain unclear.

To understand the mechanisms C. auris uses to colonize the skin, a team of researchers compared it with Candida albicans—a common skin fungus that the immune system normally clears quickly. The team used mouse models, fungal and mouse genetics, immunology, single-cell RNA-seq, and volumetric quantitative confocal microscopy. In mice, C. albicans disappeared within days, but C. auris persisted, taking refuge in hair follicles.

This work is published in Science in the paper, “The fungal pathogen Candida auris exposes chitin to trigger IFNg and persist in hair follicles.”

“Candida auris colonizes skin way better than most other fungi, setting it up to invade once the immune system is weakened,” said Dean Merrill, MD, a dermatologist and professor at UCSF. “The big clinical problem is that we have no effective way to remove it from the skin.”

The researchers discovered not only that C. auris colonized mouse skin with higher titers and greater persistence than C. albicans—exhibiting direct hair-binding activity and a strong tropism to hair follicles—but also that the two produced very different immune responses.

More specifically, they write, “Whereas C. albicans elicited a host-protective type 3/17 skin immune response driven by interleukin-17A (IL-17A), as previously described, C. auris triggered a type 1–skewed immune response, characterized by hair follicle–associated expansion of type 1 conventional dendritic cells (cDC1), type 1 cytotoxic T cells (Tc1), and T helper 1 immune cells (Th1), as well as increased interferon-γ (IFNγ) production. IFNγ signaled directly to hair follicle keratinocytes, reducing the expression of genes and downstream programs driven by IL-17A and associated with skin barrier function and antimicrobial defense.”

The team also used mice with defects in cytokine signaling to establish that IFNγ “promoted the persistence of C. auris in the epidermal niche while maintaining its classic host-protective role during deeper skin or blood infections.”

The C. auris remodels its exterior cell wall to expose more chitin, which spurs immune cells to release interferon gamma around the hair follicle. The interferon gamma blocked the skin’s antifungal defenses, including IL-17. It also slowed the natural replacement of hair follicle cells, leading to a buildup of older, damaged cells—a niche where C. auris could flourish.

“Chitin is widespread in nature, so it’s not like the human skin never encounters it, but we were surprised to see that C. auris actively uses its chitin to turn the skin into a perfect nest,” said Suzanne Noble, MD, PhD, professor of microbiology at UCSF.

The findings reveal potential targets for preventing C. auris from persisting on the skin. One approach could involve drugs that tilt the immune system away from interferon gamma signals and toward IL-17, which drives the skin’s normal antifungal clearing process. Or perhaps drugs that block chitin could prevent the fungus from amplifying the interferon gamma signals. More broadly, the researchers say the work offers a new way to think about how microbes can quietly coexist with us before becoming pathogenic.