Veit Blauhut

Claudia Teutschbein

Mathias Andersen

Manuela Brunner

Carmelo Cammalleri

Ksenija Cindrić Kalin

David Finger

Marijke Huysmans

Kiril Manevski

Marzena Osuch

Perceiving and managing the 2018 & 2019 droughts in Europe: is there a need for macro-governance in Europe?

published or not. The documents may come

Over 30 European countries joined the Panta Rhei-Drought in the Anthropocene survey. Overall, only few countries have an operational drought management strategy, whereas a majority of participants stressed the need for one.

The purpose of this survey is to analyse Europe's nation-specific perception of drought, current drought risk management, and the effects of the drought events of 2018 and 2019 (if affected). The regional and national comparison will hopefully allow us to draw conclusions to foster drought risk management and lower existing vulnerabilities. For each affected country, a national representative is in charge of the translation, distribution, and evaluation of the uniform questionnaire. This questionnaire consist of 26 questions that are concerned with how regional municipalities/organisations/companies work with the various effects of drought.

METHODS

RESULTS FROM GERMANY

CONCLUSION -Germany

Governance need ?

Germany suffered a variety of impacts in recent drought events. Germany has no common drought declaration scheme, neither are there any common practices across counties.

Germany could benefit from a Drought Risk Management Strategy implemented in the national water act?

We hypothesise that Europe would benefit from a common Drought Risk

Management strategy in order to foster the mitigation of drought impacts.

To reach this aim we have to:

  Responses by participating countries at the end of April 2020. Perceiving and managing the 2018 & 2019 droughts in Europe: is there a need for macro-governance in Europe? Veit Blauhut, Claudia Teutschbein, Mathias N. Andersen, Manuela I. Brunner, Carmelo Cammalleri, Ksenija Cindrić Kalin, David C. Finger, Marijke Huysmans, Kiril Manevski, Marzena Osuch, Renata J. Romanowicz, Kerstin Stahl, Michael Stoelzle, Anne F. Van Loon, Michelle T.H. Van Vliet, Niko Wanders, Jürgen Vogt, Jean-Philippe Vidal, and Patrick Willems veit.blauhut@hydrology.uni-freiburg.de Want to fill the white-space of your country? Contact me: veit.blauhut@hydrology.uni-freiburg.de • Germany was impacted by drought • Impacts in 2019 were perceived less severe than 2018, even though agricultural impacts increased • Impacts in 2019 were strongly influenced by the preceding year's event • Drought is expected to occur more often & severe • A more intense water management is required • Few drought classification schemes exist • Almost no proactive drought management plans exist • Only few emergency plans exist (reactive) • Main reason for the absence of drought management: • Insufficient resources • Waiting for governmental advice • Politcal ignorance, lack of EU legislation Which sectors / services / businesses were negatively affected in your municipality by the drought during the of...

  • Show the impacts of drought across Europe • Raise public and governmental awareness • Increase Science-Policy interfacing • Proof the added value by good practise examples


Storm Hassell-Hart

Sarah Picaud

Raphael Lengacher

Joshua Csucker

Regis Millet

Gilles Gasser

Roger Alberto

Hannah Maple

Robert Felix

Zbigniew J Leśnikowski

Helen J S Stewart

Timothy J Chevassut

Simon Morley

Panagis Filippakopoulos

John Spencer
email: j.spencer@sussex.ac.uk

Probing BRD Inhibition Substituent Effects in Bulky Analogues of (+)-JQ1

Keywords: Cancer, epigenetics, bioorganometallic chemistry, bromodomain, benzodiazepines

Dedicated to Prof. Antonio Togni for his seminal contributions to organic chemistry and catalysis. Thanks, Antonio, for introducing me the delights of ferrocene chemistry (JS, ETH (1994-96)) and for your continued friendship, sense of humour and mentoring

Introduction

The study of post-translational modifications (PTMs) is an area of current great scientific importance in medicine and biology, with bromodomains (BRDs) emerging as important acetylated lysine (Kac) epigenetic reader targets in medicinal chemistry. [1][2][3][4] In total, there are 61 BRDs in the "write-readerase" "epigenetic code" and inhibitors are being sought in order to elucidate their biological and clinical relevance. The BET (bromodomain and extraterminal) BRD family includes BRD-containing proteins (BRD2, BRD3, BRD4) as well as the BRD testis-specific protein (BRDT). Each protein has two tandem N-terminal bromodomains as well as an extra-terminal protein interaction domain (ET). [5][6][7][8] One well-documented chemical probe is the BET BRD4 inhibitor (+)-JQ1, which has applications in cancer, inflammation and even in contraception. [9,10] The crystal structure of (+)-JQ1 showed it to bind in the Kac pocket of BRD4(1) with the methyl-triazole unit acting as a Kac mimic. Its enantiomer (-)-JQ1 was found to be significantly less active. Recently, we disclosed the nM active (+)-JD1 (2a), ananalogue of (+)-JQ1, which has a bulky aminoferrocene moiety in place of the t-Bu ester. [START_REF] Clegg | Advancements in the Development of non-BET bromodomain chemical probes[END_REF] Its activity was lower than that of its organic analogue, typically around 3 -4-fold less active (e.g. vs. BRD4(1); (+)-JQ1 vs (+)-JD1: KD = 110 nM vs 400 nM, respectively) possibly due to the structural flexibility of the metallocene group, which was found to adopt different conformations in its protein crystal structure in BRD4(1). The presence of the ferrocene group allowed for a marginal supplementary increase in cytotoxicity by the generation of reactive oxygen species (ROS) through formation of a ferrocenium species. However, this was much less pronounced than in a previous ferrocen-containing HDAC inhibitor [12] We, therefore, wished to synthesise other metal-based analogues in order to explore structure activity relationships, steric bulk, versus bulky organic moities, where, obviously, steric bulk predominates.

Results and Discussion

Given that steric bulk appears to be tolerated around the ester unit in (+)-JQ1 analogues, we now report our findings on other series of related analogues, where, crucially, the Ot-Bu ester has been replaced by amides containing relatively large organometallic or metalloid groups. Notably, bulky, organic, "escape from flatland,' [13] , adamantylamine bioisosteres of ferrocenylamine, [14] , were also synthesized (Scheme 1) as an extension of recent work from our group, [15] , which showed similar activities of adamantyl vs ferrocenyl derivatives. The synthesis of the new analogues was achieved using known protocols. [START_REF] Clegg | Advancements in the Development of non-BET bromodomain chemical probes[END_REF] The t-Bu ester of (+)-JQ1 was treated with trifluoroacetic acid and the resulting acid (+)JQ1-OH was reacted with various amines 1a -1e using standard coupling reagents. We were able to synthesise rhenium (2b), [16] Scheme 1. Synthesis of (+)-JQ1 analogues. carborane (2c), [17][18][19] , (in the present work p-carborane isomer was used) as well as adamantyl analogues (2d, 2e) in moderate to good yields. All new compounds were characterised by 1 H, 13 C NMR spectroscopy, HRMS and analysed by HPLC purity (Scheme 1). As a positive control, we also synthesised the reported iridiumcontaining BRD4 inhibitor 3a. [20] With these compounds in hand, we tested their biochemical activity against BRDs. Thermal melt (ΔTm) data of these compounds against BET and other BRDs were obtained (Figure 2). Compared with (+)-JQ1, the p-carborane and iridium complexes 2c, 3a, respectvely, had relatively weak activity. The organic compound 2e showed better affinity toward the second bromodomains of BETs, more so than the other adamantyl analogue, 2d. Thermodynamic evaluation of binding using Isothermal titration calorimetry (ITC) of 2e against the two BRDs of BRD4 showed higher affinity for BRD4(1), however, albeit with differences in entropic contributions between the two domains (Table 1). We selected 2e for further investigation on the basis of its relatively high affinity for BRD4(2) domains as with previous studies from our group e.g. with RVX208. [21] Unfortunately, we were unable to obtain crystals of 2e in BRD4(2) or BRD2(2). Binding to the first bromodomain of BRD4 was established in a high resolution co-crystal structure (Figure 2c) confirming its Kac-competitive binding mode. The ligand adopts the classical Kac mimetic pose previously found in other thieno-and benzo-diazepine chemotypes, with the methyltriazole moity inserting into the hydrophobic recognition site and engaging the conserved asparagine (N140), while the adamantane apendix rigidly extends away from the binding site. Finally, we wished to explore the ability of 2e to suppresses c MYC and S100A8 in THP-1 cells. [START_REF] Clegg | Advancements in the Development of non-BET bromodomain chemical probes[END_REF] The acute myeloid leukaemia (AML) cell line THP-1 was treated with 1 M 2e or 1M (+)-JQ1 (positive control) for 24 h prior to RNA extraction (Figure 3). Gene expression was determined by using quantitative Real Time PCR (qPCR) and normalised using the housekeeping gene GAPDH. Both cMYC and S1008 have previously been shown to be supressed by (+)-JQ1. [22] These results showed that 2e was even more effective than (+)-JQ1.

C o n t r o l 1  M J Q 1 1  M 2

Conclusions

BRD inhibitors based on (+)-JQ1 and (+)-JD1, including rhenium and carborane analogues, with bulky amide groups have been synthesized and characterized. The related, sterically bulky, adamantyl analogue (2e) displayed excellent binding (KD = ca. 130 nM) to BRD4(1) with a ca. two-fold selectivity over BRD4(2) (KD = ca. 260 nM) and its crystal structure in the first bromodomain of BRD4 was determined. use of 2e, which also suppresses c MYC in THP-1 cells, as a tool compound in cell-based systems, is currently underway in our laboratory and will be disclosed in due course.

Experimental Section

Synthetic chemistry details, including NMR, mass spectrometry, HPLC, are as reported recently. [START_REF] Clegg | Advancements in the Development of non-BET bromodomain chemical probes[END_REF] Synthetic Chemistry (+)-JQ-1-OH. A mixture of (+)-JQ-1 (200 mg, 0.438 mmol) and anhydrous DCM (50 mL) was cooled to 0 °C. To the mixture was added dropwise trifluoroacetic acid (20 mL) and the resulting mixture was warmed to ambient temperature and stirred for 16 h under an argon atmosphere. The reaction was concentrated under reduced pressure and to the residue was added a 1:1 mixture of DCM:Et2O. The solvent was removed under reduced pressure and the sequence repeated five times to give (+)-JQ-1-OH as a yellow solid of sufficient purity to be utilised in subsequent reactions without further manipulation.

Rhenium Analogue of (+)-JQ-1 (2b)

To (+)-JQ-1-OH (11.3 mg, 0.0283 mmol) was added anhydrous THF (1.25 mL) and anhydrous DIPEA (9.9 µL, 0.0566 mmol) under an argon atmosphere. To the resulting mixture was added HBTU (21.5 mg, 0.0566 mmol) and rhenium complex 1b (15.6 mg, 0.0283 mmol). The resulting mixture was heated to 40 °C and stirred for 72 h under an argon atmosphere. The reaction was cooled to ambient temperature and to the mixture was added DCM (5 mL) and a solution of 2M aqueous NaOH (5 mL). The resulting biphasic mixture was separated and the resulting organic extract washed with a solution of 2M HCl (5 mL) followed by brine (5 mL), dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to give an orange gum (59 mg). The resulting residue was purified by automated flash column chromatography (EtOAc/MeOH, 100:0 -80:20, 4 g SiO2). The appropriate fractions were combined and concentrated under reduced pressure to give the Rhenium analogue of JQ-1 as a yellow solid (9 mg, 38% over 2 steps

Carborane Analogue of JQ-1 (2c)

To (+)-JQ-1-OH (44 mg, 0.110 mmol) was added anhydrous THF (5 mL) and anhydrous DIPEA (38 µL, 0.219 mmol) under an argon atmosphere. To the resulting mixture was added HBTU (83 mg, 0.219 mmol) and (aminoopropyl-p-carborane), [START_REF] Bednarska | Effect of adenosine modified with a boron cluster pharmacophore on reactive oxygen species production by human neutrophils[END_REF] , (52 mg, 0.219 mmol). and the mixture stirred at ambient temperature for 16 h under an argon atmosphere.To the mixture was added DCM (10 mL) and a solution of 2M aqueous NaOH (10 mL). The resulting biphasic mixture was separated and the resulting organic extract washed with a solution of 2M HCl (10 mL) followed by brine (10 mL), dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to give a yellow solid (94 mg). The resulting residue was purified by automated flash column chromatography (EtOAc/MeOH, 100:0 -70:30, 12 g SiO2). The appropriate fractions were combined and concentrated under reduced pressure to give 2c as a white solid (72 mg, 95% over two steps). LCMS (UV, ESI) Rt = 25.90 min, [M-H] + m/z = 585.2, 95% purity. 1 H NMR (600 MHz, d6 -DMSO): δ = 8.17 (1H, t, J = 5.8 Hz), 7.51-7.47 (2H, m), 7.44-7.40 (2H, m), 4.46 (1H, dd, J = 8.5, 5.7 Hz), 3.40-3.31 (11H, m), 3.20 (1H, dd, J = 14.9, 5.7 Hz), 3.11 (1H, dd, J = 14.9, 5.7 Hz),

N-((1s,3R)-Adamantan-1-yl)-2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide (2d).

To (+)-JQ-1-OH (16 mg, 0.04 mmol) was added anhydrous THF (2 mL) and anhydrous DIPEA (14 µL, 0.08 mmol) under an argon atmosphere. To the resulting mixture was added HBTU (30 mg, 0.08 mmol) and 1-adamantanamine (12 mg, 0.08 mmol) and the mixture stirred at ambient temperature for 16 h under an argon atmosphere. To the mixture was added DCM (5 mL) and a solution of 2M aqueous NaOH (5 mL). The resulting biphasic mixture was separated and the resulting organic extract washed with a solution of 2M HCl (5 mL) followed by brine (5 mL), dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to give a yellow solid (40 mg). The resulting residue was purified by automated flash column chromatography (EtOAc/MeOH, 100:0 -70:30, 40 g SiO2). The appropriate fractions were combined and concentrated under reduced pressure to give the title compound as a white solid (16 mg, 75% over two steps

N-((1s,3R)-Adamantan-1-ylmethyl)-2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide (2e).

To (+)-JQ-1-OH (16 mg, 0.04 mmol) was added anhydrous THF (2 mL) and anhydrous DIPEA (14 µL, 0.08 mmol) under an argon atmosphere. To the resulting mixture was added HBTU (30 mg, 0.08 mmol) and 1-adamantanemethylamine (14 µL, 0.08 mmol) and the mixture stirred at ambient temperature for 16 h under an argon atmosphere. To the mixture was added DCM (5 mL) and a solution of 2M aqueous NaOH (5 mL). The resulting biphasic mixture was separated and the resulting organic extract washed with a solution of 2M HCl (5 mL) followed by brine (5 mL), dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to give a yellow solid (49 mg). The resulting residue was purified by automated flash column chromatography (EtOAc/MeOH, 100:0 -70:30, 4 g SiO2). The appropriate fractions were combined and concentrated under reduced pressure to give the title compound as a white solid (18 mg, 82% over two steps

Iridium (III) Complex (3a)

To a mixture of dichlorotetrakis(2-(2-pyridinyl)phenyl)diiridium(III) (25 mg, 0.023 mmol) and anhydrous acetonitrile (2.5 mL) was added silver triflate (12 mg, 0.046 mmol) and the resulting mixture stirred at ambient temperature for 16 h under an argon atmosphere. The resulting mixture was filtered through celite®, washing with Et2O. The resulting solution was concentrated under reduced pressure to give a yellow solid (32 mg, 95%). Data are in agreement with those previously reported. [20]

Figure 1 .

 1 Figure 1. Iridium-containing positive control (3a).

Figure 2 .

 2 Figure 2. In vitro evaluation of compound 2 series. A) Thermal melt (ΔTm) data of 2b-e against BET and other BRDs using proteins at a final concentration of 2 μM and ligands at 10 μM. B) Thermodynamic evaluation of 2e binding to BRD4 bromodomains, by isothermal titration calorimetry. Raw injection heats for titrations of BRD4(1) or BRD4(2) into solutions of 2e are shown in the left panels (reverse titrations). The right panel shows the normalized binding enthalpies corrected for the heat of protein dilution as a function of binding site saturation (symbols as indicated in the figure). Solid lines represent a nonlinear least squares fit using a single-site binding model. Titrations were performed in 50 mM HEPES pH 7.5 (at 25°C), 150 mM NaCl and 15°C while stirring at 750 rpm. (BRD4(1): KD = 133 ± 6 nM; BRD4(2): KD = 257 ± 21 nM). C) Crystal structure of 2e bound onto BRD4(1). (PDB: 7AJN). Key residues of the Kac-binding pocket are annotated and conserved structural waters are shown. D) The ligand (2e) 2Fc-Fo map contoured at 2σ is shown in detail.

Figure 3 .

 3 Figure 3. Comparison of effects of 2e vs (+)-JQ1 in suppressing c MYC and S100A8 (triplicate data).

Table 1 -

 1 Isothermal Titration Calorimetry of human BRD4 bromodomains with compound 2e. Titration was carried out in 50 mM HEPES pH 7.5 (at 25 °C), 150 mM NaCl and 15 °C while stirring at 750 rpm. The protein was titrated into the compound solution (reverse titration).

	Protein	Ligand	[P] (μM)	[L] (μM)	KD (nM)	N	ΔH obs (kcal/mol)	TΔS (kcal/mol)	ΔG (kcal/mol)
	BRD4(1)	2e	550	22	133 ± 6	0.96 ± 0.002	-10.20 ± 0.03	-1.135	-9.07
	BRD4(2)	2e	586	25	257 ± 21	0.97 ± 0.004	-6.53 ± 0.05	2.154	-8.69

  ). LCMS (UV, ESI) Rt = 24.64 min, [M-H] + m/z = 548.2, 96% purity. 1 H NMR (600 MHz, CDCl3): δ = 7.41 (2H, d, J = 8.2 Hz), 7.35-7.29 (2H, m), 6.50 (1H, t, J = 6.4 Hz), 4.60 (1H, dd, J = 7.4, 6.4 Hz), 3.60 (1H, dd, J = 14.1, 7.4 Hz), 3.35 (1H, dd, J = 14.1 Hz, 6.5 Hz), 3.01 (1H, dd, J = 13.4, 6.4 Hz), 2.95 (1H, dd, J = 13.4, 6.4 Hz), 2.67 (3H, s), 2.39 (3H, s), 1.97-1.92 (3H, m), 1.71-1.67 (3H, m), 1.66 (3H, s), 1.61-1.56 (3H, m), 1.50-1.46 (3H, m), 1.46-1.41 (3H, m).13 C NMR (151 MHz, CDCl3): δ = 170.7, 164.0, 155.9, 150.0, 136.9, 136.7, 132.2, 131.1, 131.0, 130.7, 130.0, 128.8, 54.8, 51.2, 40.3, 39.8, 37.0, 33.9, 28.3, 14.5, 13.2, 12.0. HRMS (ESI-[+H]) m/z: Calcd for C30H34ClN5OS 548.2251; Found 548.2231.

Acknowledgements. EPSRC (EP/P026990/1, S.H.-H., J.S.), MRC (MR/N010051/1, P.F.) and NSC Poland (2014/13/B/NZ1/03989, ZJL) are thanked for funding. The authors thank the Diamond Light Source for beam time (proposal mx19301) and the staff of beamlines I03 for assistance with crystal testing and data collection. The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada, Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD grant no. 115766], Janssen, Merck KgaA Darmstadt Germany, MSD, Novartis Pharma AG, Ontario Ministry of Economic Development and Innovation, Pfizer, FAPDF, CAPES, CNPq, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome [106169/ZZ14/Z]. This work was funded by the Swiss National Science Foundation (Grant Sinergia CRSII5_173718) and has received support under the program «Investissements d'Avenir » launched by the French Government and implemented by the ANR with the reference ANR-10-IDEX-0001-02 PSL (G.G.).

Supplementary Material

Supporting information for this article is available on the WWW under http://dx.doi.org/10.1002/MS-number.

Abbreviations. HBTU (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl uronium hexafluorophosphate. DIPEA: N,N-Diisopropylethylamine.

Author Contribution Statement

J.S., P.F. designed the project and wrote the manuscript with writing and editing help from G.G., R.M., H.M., R.F. and S.M. S.H-H synthesized the final compounds and intermediates with help from J.C., R.L., Z.J. L. and R. A.. S.P., T. J. C., H.J.S.S. and P.F. performed biological evaluation of the final compounds as well as x-ray structural analysis.

Entry for the Table of Contents

Twitter (+)-JQ1 analogues containing bulky groups show good BRD4 affinity especially the adamantyl analogue 2e. @GasserGroup, @UZH_Chemistry, @ChemistrySussex; @JSpencerlab


Jeremie Bonneau

Flora Branger

Helene Castebrunet

Gislain Lipeme Kouyi

Urbanisation, Climate Change and Stormwater Management Strategies: Modelling the Future Flow Regimes of Peri-Urban Streams in Lyon, France

The health of peri-urban rivers is under pressure due to accelerating urbanisation and climate change. Predicting their flow regime in a few decades is a challenge. We modelled the hydrology of two peri-urban catchments with the physically-based, spatially distributed hydrological model J2000P at the hourly timestep. We aimed to quantify the respective impact of 1/ real life scenarios of future urban development, 2/ climate change and 3/ potential stormwater management strategies ('disconnecting imperviousness') on the flow regimes of both rivers. We developed J2000P to integrate a site-scale model of green infrastructure. For several scenarios (combination of land use, climate, stormwater management scenarios), we looked at a range of hydrological signatures and metrics such as catchment water balance, baseflow behaviour, storm related metrics, combined-sewer overflows to calibrate the model and quantify the relative impact of landuse change, climate change and stormwater management. Preliminary results show that the model can be well calibrated to reproduce current flow regimes. Impending simulations will take into account future urbanisation scenarios and climate change characteristics, as well as disconnection strategies. Results will inform local public policies about the extent of green infrastructure implementation required to keep healthy rivers in the coming decades.

INTRODUCTION AND GOAL

The health of peri-urban rivers is under pressure due to accelerating urbanisation and climate change. Predicting their flow regime in a few decades is a challenge. We aim to model the hydrology of a peri-urban catchment located in the western suburbs of Lyon, France. We aim to study the response of the flow regime to:

1/ future urbanisation 2/ climate change 3/ stormwater management scenarios, especially the implementation of green infrastructure that aims to mimic the natural water fluxes (infiltration, evapotranspiration) In this poster, we report the development of the model, the construction of scenarios and results of the calibration of the model.

CATCHMENT CHARACTERISTICS

The catchment is located in the western suburbs of Lyon, France. It is 20 km , and drains mixed landuse: ranging from forested areas upslope, farmland and urban areas downslope of the catchment (see Figure below).

The catchent is a sub-catchment of the Yzeron catchment, part of the long term observatory in Urban Hydrology OTHU (http://www.graie.org/othu/ (http://www.graie.org/othu/)) From 1997 to 2019, the catchment averaged 716 mm of rainfall with an average potential evapotranspiration of 762 mm. The catchment is drained by a stream (le Ratier), that has been monitored for 10 years now. Catchment geology is uniform (metamorphic rocks), and soils are a range of shallow sandy-loams (1-3 m in average).

Stormwater in urban areas are drained by a combiend stormwater/sewer network, meaning that most rainfall on impervious areas is collected and drained out of the catchment (except for occasional combined sewer overflow spills).

PRESENTATION AND DEVELOPMENT OF THE MODEL

Model used

In order to model the catchment and its stream, we used the hydrological model J2000P [START_REF] Labbas | Modélisation hydrologique de bassins versants périurbains et influence de l'occupation du sol et de la gestion des eaux pluviales: Application au bassin de l'Yzeron (130km2)[END_REF], a distributed, process-based model, a variant of J2000, developed by the University of Jena in Germany [START_REF] Krause | Quantifying the impact of land use changes on the water balance of large catchments using the J2000 model[END_REF], adapted to periurban catchment. The model runs at the hourly timestep.

The catchment was discretised in Hydrological Response Units (HRU), homogeneous in landuse, soil cover and underlying geology (Figure below). Each HRU is thus homogeneous hydrologically.

Within each HRU, J2000P takes into account the following hydrological processes: interception of rainfall by the vegetation, impervious and surface runoff, infiltration into a 'medium pore' soil storage from which evapotranspiration is drawn and into a 'large pore' soil storage which produce subsurface interflow and percolation from the soil to a groundwater store that produce baseflow. Flow is therefore divided into thrree components : surface runoff, interflow, baseflow. HRUs are connected to each other or to river reaches through topology. The model takes into account multiple drainage networks: natural river networks as well drainage/sewer networks with outlets out of the catchment; into the river networks and through sewer overflow devices.

Input data

Rainfall data and flow data were source from the Hydrology Observatory Database Yzeron (Base de Données des Observatoires en Hydrologie, BDOH, https://bdoh.irstea.fr/ (https://bdoh.irstea.fr/)).

Development of a green infrasructure model

We developed a Green Infrastructure module into the model to able to take into account systems such as infiltration trenches, green roofs, etc. Some of the surface runoff is diverted to green infrastructures (GI), that is either 1/ stored within the GI 2/ evapotranspired 3/ infiltrated into the native soil or 4/ connected back to the network.

The Figure below presents a simplified conceptual diagram of the model with the addition of a green infrastructure module.

Future urbanisation scenarios

The catchment imperviousness was around 12% in 2008. The catchment is undergoing urbanisation, with suburbs covering farmland. Following the work of Dodanne (2014), maps of future urbanisation were sourced (Figure below). Based on an expert panel judgment, local stakeholders, demographic trends and regulations projections, the scenarios for future landuse in 2030 were as realistic as possible. Several scenarios were considered: current sprawl trend, acceleration of the trend, densification to limit the urban sprawl. We discretized these maps to be used as input of the model.

Future stormwater management scenarios

We are also setting up a range of stormwater management scenarios, with different degrees of 'disconnection' of newly impervious areas as well as some retrofit management scenarios in existing impervious areas. We estimated where it was possible and plausible to set up infiltration trenches, bioretention basins, green roofs, porous pavement, to manage runoff generated by impervious areas.

CLIMATE SCENARIOS

We aim to study the vulnerability of the catchment to future climate change. To do so, hourly timeseries of precipitation are needed as input data for the model. Future climate projections are for the catchment through the French portal DRIAS (http://www.drias-climat.fr/), based on different greenhouse gases emissions and representative concentration pathways. However, these data are not directly usable in this study, since they are only available at the monthly or daily timestep, too coarse for a fast, responsive urban catchment. In addition, climate projections fail to adequately represent convective meteorological processes which are important for subdaily storms and rainfall patterns.

Therefore, we chose a bottom-up approach [START_REF] Sauquet | Water restrictions under climate change: a Rhône-Mediterranean perspective combining bottom-up and top-down approaches[END_REF], also called approach 'by perturbation' to study the vlnerability of the catchment to climate change. We transformed present hourly timeseries (1990-2020) of rainfall and evapotranspiration by applying perturbation factors, to obtain future, hourly timeseries. The concept of the bottom-up approach is illustrated below:

Evapotranspiration

Present evapotranspiration was transformed into six future evapotranspiration timeseries through six perturbations in the temperature regime, applied to the evapotranspiration timeseries with the Oudin formula [START_REF] Oudin | Which Potential Evapotranspiration Input for a Lumped Rainfall-runoff Model?[END_REF]:

With ETP* the new, perturbed evapotranspiration timeseries, ETP the current evapotranspiration timeseries, ΔT the monthly change in temperature, and Ra the catchment radiation (W/m2).

The change in monthly temperature ΔT (six scenarios) is modeled as a sine curve with varying annual mean increase +1, 3, 5 °C, annual amplitude of 0.5 or 1.5 °C, with a maximum change in August, as shown in the following Figure:

Rainfall

To create rainfall perturbation factors, we plotted probability of exceedance curves for daily rainfall for the catchment ("Pollionay") and for the southern French city of Orange (it is observed the climate in south easy France is drifting from south to North, with precipitation and temperatures in Lyon now looking like historical records of cities 50 km south). The perturbation factor was considered the quantile by quantile ratio, as: with p daily precipitation corresponding to the quantile q. Four perturbation factors were calculated as multiples of the initial perturbation factor calculated with Orange data (illustrated below). With these four perturbations, we take into account several possible increases of high rainfall and several possible changes in low rainfall patterns. These perturbations factors, calculated thanks to daily timeseries, are then applied to hourly present timeseries to obtain a range of possible future rainfall timeseries, to be used as input for the hydrological model.

AUTHOR INFORMATION