Biology subjects

de Haan, S.

Publications and source records attributed to de Haan, S..

8 recordsLinked to original sources

bioRxiv2026

Biocatalytic Production of Galantamine in Yeast through Cytochrome P450 Optimization

Galantamine is a pharmaceutically relevant Amaryllidaceae alkaloid used for the treatment of Alzheimer's disease. Its structural complexity and low abundance in plants motivate the development of alternative manufacturing routes. Here, we established the first engineered yeast platform for the biocatalytic production of galantamine from 4OMe-norbelladine. Heterologous expression of the downstream pathway enzymes NtCYP96T6, NtNMT1, and NtAKR1 from Narcissus cv. Tete-a-Tete in Saccharomyces cerevisiae was complemented by optimization of cultivation temperature, carbon source, and medium pH enabling the first demonstration of galantamine production in yeast. Systematic screening of cytochrome P450 reductase and cytochrome b partners to boost NtCYP96T6 activity led to the identification of a novel reductase mined from the Narcissus pseudonarcissus transcriptome, NpCPR, which supported the highest pathway flux and yielded 7.0 {+/-} 0.6 mg/L galantamine, corresponding to a 7.6 % molar yield from 250 M 4OMe-norbelladine and an approximately 173-fold improvement over the parental strain. We further exploited this yeast cell factory for the precursor-directed biosynthesis of 7F-galantamine, highlighting the potential of pathway enzyme promiscuity to access new-to-nature GAL analogues that may be challenging to produce through conventional chemical synthesis. Together, this work establishes a foundation for microbial galantamine production and biosynthetic diversification of its pharmaceutically relevant scaffold.

synthetic biology↗

bioRxiv2026

Engineering Biosensors to Enhance Monoterpene Indole Alkaloid Production in Yeast

Monoterpene Indole Alkaloids (MIAs) are a diverse family of plant natural products with various medicinal applications. Although MIAs, such as vinblastine and reserpine, are clinically validated, sourcing of MIAs for clinical use or drug discovery from natural resources or via chemical synthesis is hampered due to their scarcity and chemical complexity. Refactoring MIA biosynthesis pathways in microbial cell factories could offer an alternative, more stable and potentially sustainable manufacturing route for alkaloid medicines and novel therapies. However, reaching commercially attractive titers, rates and yields remains challenging owing to the length and complexity of these metabolic pathways. One critical bottleneck is the low screening throughput and very high cost of the analytical methods used to quantify MIA for optimizing production. In this study, we evolved RamR, a promiscuous bacterial transcription factor to respond to five different MIAs, resulting in highly sensitive and selective sensor variants (EC50<10 M). X-ray crystallography and computational modeling provided insight into the MIA binding of the evolved biosensor variants. The RamR biosensing platform was functionalized in yeast and subsequently applied in a cost-effective semi-throughput screening campaign of a 188-gene overexpression library to identify high-performing cell factory designs for strictosidine, the common precursor for all MIAs. The fluorescent biosensor signal correlated with HPLC quantification (r2 = 0.932) allowing identification of single metabolic engineering hits which when combined yielded a maximum titer of >220 mg/L strictosidine, 3-fold higher than the parental reference strain. This study demonstrates the development of selective biosensors for MIAs and the cost-effective identification of novel metabolic engineering hits for optimizing MIA production in microbial cell factories.

synthetic biology↗

bioRxiv2025

Mechanisms of resistance to VHL loss-induced genetic and pharmacological vulnerabilities

The von Hippel-Lindau tumor suppressor (VHL) is a component of a ubiquitin ligase complex that normally controls cellular responses to hypoxia. Endogenous VHL is also utilized by proteolysis-targeting chimera (PROTAC) protein degraders, a promising class of anti-cancer agents. VHL is broadly essential for cell proliferation, yet it is a key tumor suppressor in renal cell carcinoma. To understand the functional consequences of VHL loss, and to identify targeted approaches for the elimination of VHL null cells, we have used genome-wide CRISPR-Cas9 screening in human renal epithelial cells. We find that, upon VHL loss, the HIF1A/ARNT complex is the central inhibitor of cellular fitness, suppressing mitochondrial respiration, and that VHL null cells show HIF1A-dependent molecular vulnerabilities that can be targeted pharmacologically. Combined VHL/HIF1A inactivation in breast and esophageal cancer cells can also provide resistance to ARV-771, a VHL-based bromodomain degrader that has anti-cancer activity. HIF1A stabilization can thus provide opportunities for early intervention in neoplastic VHL clones, and the VHL-HIF1A axis may be relevant for the development of resistance to the emerging class of PROTAC-based cancer therapies.

cancer biology↗

bioRxiv2024

Hierarchical Target Learning in the Mammalian Neocortex: A Pyramidal Neuron Perspective

1Computational neuroscience currently debates two competing hypotheses to explain hierarchical learning in the neocortex: deep learning inspired approximations of the backpropagation algorithm, where neurons adjust synapses to minimize an error, and target learning algorithms, where neurons learn by reducing the feedback needed to achieve a desired target activity. While both hypotheses have been supported by theoretical studies, there is currently no empirical test that compares them directly. Here we provide such tests in the mouse neocortex by evaluating both hypotheses against experimental data at the single cell level and at the population level. At the single cell level, we conduct in vitro experiments that clarify the relationship between algorithmic learning signals and synaptic plasticity within individual pyramidal neurons. At the population level, we analyze in vivo calcium imaging data in the lateral visual cortex. By combining in vivo and in vitro data we reveal a critical discrepancy between neocortical hierarchical learning and canonical machine learning.

neuroscience↗

bioRxiv2024

Decision support for managing an invasive pathogen through efficient clean seed systems: Cassava mosaic disease in Southeast Asia

CONTEXTEffective seed systems must both distribute high-performing varieties efficiently and slow or stop the spread of pathogens and pests. Epidemics increasingly threaten crops around the world, endangering the incomes and livelihoods of smallholder farmers. Responding to these food and economic security challenges requires stakeholders to act quickly and decisively during the early stages of invasions, typically with very limited resources. The recent introduction of cassava mosaic virus into southeast Asia threatens cassava production in the region. OBJECTIVESOur goal in this study is to provide a decision-support framework for efficient management of healthy seed systems, applied to cassava mosaic disease. The specific objectives are to (1) evaluate disease risk in disease-free parts of Cambodia, Lao PDR, Thailand, and Vietnam by integrating disease occurrence, climate, topology, and land use, using machine learning; (2) incorporate this predicted environmental risk with seed exchange survey data and whitefly spread in the landscape to model epidemic spread in a network meta-population model; and (3) use scenario analysis to identify candidate regions to be prioritized in seed system management. RESULTS AND CONCLUSIONSThe analyses allow stakeholders to evaluate strategy options for allocating their resources in the field, guiding the implementation of seed system programs and responses. Fixed rather than adaptive deployment of clean seed produced more favorable outcomes in this model, as did prioritization of a higher number of districts through the deployment of smaller volumes of clean seed. SIGNIFICANCEThe decision-support framework presented here can be applied widely to seed systems challenged by the dual goals of distributing seed efficiently and reducing disease risk. Data-driven approaches support evidence-based identification of optimized surveillance and mitigation areas in an iterative fashion, providing guidance early in an epidemic, and revising them as data accrue over time.

ecology↗

bioRxiv2024

Jag1 represses Notch activation in lateral supporting cells and inhibits an outer hair cell fate in the medial compartment of the developing cochlea

Notch signaling regulates both inner and middle ear morphogenesis and establishes a strict pattern of sensory cells in the organ of Corti in the mammalian cochlea. Patients with Alagille syndrome have impaired Notch signaling ([~]94% with JAG1 mutations) resulting in sensorineural and conductive hearing loss. Here, we investigate the function of Jag1-mediated Notch activation in cochlear patterning and signaling using the Jag1 "Nodder" (Jag1Ndr/Ndr) mouse model of Alagille syndrome. Jag1Ndr/Ndr mice exhibited severe vestibular and auditory deficits and a dose-dependent increase in ectopic inner hair cells and a reduction in outer hair cells. Single cell RNA sequencing of the organ of Corti demonstrated a global dysregulation of genes associated with inner ear development and deafness. Analysis of individual cell types indicated a novel role for Jag1 in repressing Notch activation in lateral supporting cells and revealed a function for Jag1 in gene regulation and development of outer hair cells. Additionally, "outer hair cell-like" SLC26A5 (Prestin) positive cells were present in the medial compartment and pillar cell region of Jag1Ndr/Ndr mice and exhibited location-dependent expression of the inner hair cell fate-regulator Tbx2, revisiting the potency of Tbx2 in driving inner hair cell commitment in "outer hair cell-like" cells in the Jag1-mutant IHC compartment. Our data reveals a novel role for Jag1 in repressing Notch activation in lateral supporting cells and highlights involvement for Notch signaling in inner versus outer hair cell specification and positioning.

developmental biology↗

bioRxiv2023

Napping and circadian sleep-wake regulation during healthy aging.

Study objectivesDaytime napping is frequently reported among the older population and has attracted increasing attention due to its association with multiple health conditions. Here, we tested whether napping in the aged is associated with altered circadian regulation of sleep, sleepiness and vigilance performance. MethodsSixty healthy older individuals (mean age: 69y., 39 women) were recruited with respect to their napping habits (30 nappers, 30 non-nappers). All participants underwent an in-lab 40-h multiple nap protocol (10 cycles of 80 mins of sleep opportunity alternating with 160 mins of wakefulness), preceded and followed by a baseline and recovery sleep period. Saliva samples for melatonin assessment, sleepiness and vigilance performance were collected during wakefulness and electrophysiological data were recorded to derive sleep parameters during scheduled sleep opportunities. ResultsThe circadian amplitude of melatonin secretion was reduced in nappers, compared to non-nappers. Furthermore, nappers were characterized by higher sleep efficiencies and REM sleep proportion during day-compared to night-time naps. The nap group also presented altered modulation in sleepiness and vigilance performance at specific circadian phases. DiscussionOur data indicate that napping is associated with an altered circadian sleep-wake propensity rhythm and thereby contribute to the understanding of the biological correlates underlying napping and/or sleep-wake cycle fragmentation during healthy aging. Altered circadian sleep-wake promotion can lead to a less distinct allocation of sleep into night-time and/or a reduced wakefulness drive during the day, thereby potentially triggering the need to sleep at adverse circadian phase. SIGNIFICANCE STATEMENTWhile napping has raised increasing interest as a health risk factor in epidemiological studies, its underlying regulation processes in the aged remain largely elusive. Here we assessed whether napping in the older population is associated with physiological and behavioral changes in circadian sleep-wake characteristics. Our data indicate that, concomitant to a reduced circadian amplitude in melatonin secretion, healthy older nappers are characterized by reduced day-night differences in sleep efficiency and more particularly in REM sleep, compared to their non-napping counterparts. These results suggest altered circadian response as a cause or consequence of chronic napping in the aged and thereby contribute to the understanding of nap regulation during healthy aging.

neuroscience↗

bioRxiv2019

The land-use dynamics of potato agrobiodiversity in the highlands of central Peru: a case study of spatial-temporal management across farming landscapes

In the high Andes, environmental and socio-economic drivers are transforming land use and presumably affecting the in situ conservation of potato (Solanum spp.). To monitor the use and conservation of intraspecific diversity, systematic and comparative studies across land-use systems are needed. We investigated the spatial-temporal dynamics of potato in two contrasting landscapes of Perus central Andes: a highland plateau (Huancavelica) vs. an eastern slope (Pasco). We examined household-level areal allocations, altitudinal distribution, sectoral fallowing practices, and the conservation status for three main cultivar groups: (i) bred varieties, (ii) floury landraces, and (iii) bitter landraces. Mixed methods were used to survey 323 households and the 1,101 potato fields they managed in 2012-2013. We compared the contemporary altitudinal distribution of landraces with 1975-1985 altimeter genebank data from the International Potato Center. We show that intensification occurs in each landscape through adaptations of traditional management practices while maintaining high intraspecific diversity. Access to land and production end use (sale vs. consumption) significantly affected smallholder management and differentiated the landscapes. Total areas in Huancavelica and Pasco were allocated to 82.9% vs. 74.2% floury landraces, 9.2% vs. 25.7% bred varieties, and 7.9% vs. 0.1% bitter landraces. In market-oriented Pasco, fields in sectoral fallows between 3,901 m and 4,116 m above sea level consistently contained the highest levels of landrace diversity. The bulk of diversity in subsistence-oriented Huancavelica occurred between 3,909 m and 4,324 m outside sectoral fallows. Most of the unique landraces documented were scarce across households: 45.4% and 61.7% respectively in Huancavelica and Pasco. Bred varieties showed the widest (1,100 m) and bitter landraces the narrowest (400 m) altitudinal distributions. Potato cultivation has moved upward by an average of 306 m since 1975. Landrace diversity is versatile but unevenly distributed across landscapes. This requires adaptive ways to incentivize in situ conservation.

ecology↗