Biology subjects

Lemke, C.

Publications and source records attributed to Lemke, C..

5 recordsLinked to original sources

bioRxiv2023

In vitro validated antibody design against multiple therapeutic antigens using generative inverse folding

Deep learning approaches have demonstrated the ability to design protein sequences given backbone structures [1, 2, 3, 4, 5]. While these approaches have been applied in silico to designing antibody complementarity-determining regions (CDRs), they have yet to be validated in vitro for designing antibody binders, which is the true measure of success for antibody design. Here we describe IgDesign, a deep learning method for antibody CDR design, and demonstrate its robustness with successful binder design for 8 therapeutic antigens. The model is tasked with designing heavy chain CDR3 (HCDR3) or all three heavy chain CDRs (HCDR123) using native backbone structures of antibody-antigen complexes, along with the antigen and antibody framework (FWR) sequences as context. For each of the 8 antigens, we design 100 HCDR3s and 100 HCDR123s, scaffold them into the native antibodys variable region, and screen them for binding against the antigen using surface plasmon resonance (SPR). As a baseline, we screen 100 HCDR3s taken from the models training set and paired with the native HCDR1 and HCDR2. We observe that both HCDR3 design and HCDR123 design outperform this HCDR3-only baseline. IgDesign is the first experimentally validated antibody inverse folding model. It can design antibody binders to multiple therapeutic antigens with high success rates and, in some cases, improved affinities over clinically validated reference antibodies. Antibody inverse folding has applications to both de novo antibody design and lead optimization, making IgDesign a valuable tool for accelerating drug development and enabling therapeutic design. The data generated in this study serve as a useful benchmark of diverse antibody-antigen interactions. We use this data to benchmark self-consistency RMSD (scRMSD), using ABodyBuilder2 [6], ABodyBuilder3 [7], and ESMFold [8], as a metric for assessing binding. We open source the code for IgDesign and the SPR datasets. Code and datasets can be found at https://github.com/AbSciBio/igdesign.

synthetic biology↗

bioRxiv2021

Natural variation further increases resilience of sorghum bred for chronically drought-prone environments

Climate-change-associated shifts in rainfall distribution together with a looming worldwide water crisis make drought resilience of central importance to food security. Even for relatively drought resilient crops such as sorghum, moisture stress is nonetheless one of the major constraints for production. Here, we explore the potential to use natural genetic variation to build on the inherent drought tolerance of an elite cultivar (Teshale) bred for Ethiopian conditions including chronic drought, evaluating a backcross nested-association mapping (BC-NAM) population using 12 diverse founder lines crossed with Teshale under three drought-prone environments in Ethiopia. All twelve populations averaged higher head exsertion and lower leaf senescence than the recurrent parent in the two highest-stress environments, reflecting new drought resilience mechanisms from the donors. A total of 154 QTLs were detected for eight drought responsive traits - the validity of these were supported in that 100 (64.9%) overlapped with QTLs previously detected for the same traits, concentrated in regions previously associated with stay-green traits as well as the flowering regulator Ma6 and drought resistant gene P5CS2. Allele effects show that some favorable alleles are already present in the Ethiopian cultivar, however the exotic donors offer rich scope for increasing drought resilience. Using model-selected SNPs associated with eight traits in this study and three in a companion study, phenotypic prediction accuracies for grain yield were equivalent to genome-wide SNPs and were significantly better than random SNPs, indicating that these studied traits are predictive of sorghum grain yield. Rich scope for improving drought resilience even in cultivars bred for drought-prone regions, together with phenotypic prediction accuracy for grain yield, provides a foundation to enhance food security in drought-prone areas like the African Sahel.

genetics↗

bioRxiv2021

New adaptive peaks for crops - an example from improvement of drought-resilience of sorghum in Ethiopia

O_LIAs the center of diversity for sorghum [Sorghum bicolor (L.) Moench], elite cultivars selected in Ethiopia are of central importance to sub-Saharan food security. Despite being presumably well adapted to their center of diversity, elite Ethiopian sorghums nonetheless experience constraints to productivity, for example associated with shifting rainfall patterns associated with climate change. C_LIO_LIA sorghum backcross nested association mapping (BC-NAM) population developed by crossing thirteen diverse lines pre-identified to have various drought resilience mechanisms, with an Ethiopian elite cultivar, Teshale, was tested under three rain-fed environments in Ethiopia. C_LIO_LI27, 15, and 15 QTLs with predominantly small additive effects were identified for days to flowering, days to maturity, and plant height, respectively. Many associations detected in this study corresponded closely to known or candidate genes or previously mapped QTLs, supporting their validity. Field tests show drought resilience to be improved by incorporation of adaptations from the diverse donor lines. C_LIO_LIThe expectation that genotypes such as Teshale from near the center of diversity tend to have a history of strong balancing selection, with novel variations more likely to persist in small marginal populations, was strongly supported in that for these three traits, nearly equal numbers of alleles from the donor lines conferred increases and decreases in phenotype relative to the Teshale allele. Such rich variation provides a foundation for selection to traverse a valley of reduced yield and arrive at a new adaptive peak, exemplifying the nature of efforts that may be necessary to adapt many crops to new climate extremes. C_LI Societal Impact StatementIn Ethiopia, agriculture is the largest economic sector and contributes 48% of the nations GDP, and sorghum provides more than one third of the cereal diet and is widely grown for food, feed, brewing, and construction purposes. With a worldwide water crisis looming, developing drought tolerant sorghum is vital in rain-fed environments, particularly in sub-Saharan Africa. Addressing such issues often requires a far-reaching approach to identify and incorporate new traits into a gene pool, followed by a period of selection to re-establish an overall adaptive phenotype. This study revealed that with the enormous altitudinal variation of a country such as Ethiopia, somewhat different lines may be needed for different locales.

genetics↗

bioRxiv2019

Mapping the S1 and S1 subsites of cysteine proteases with new dipeptidyl nitrile inhibitors as trypanocidal agents

The cysteine protease cruzipain is considered to be a validated target for therapeutic intervention in the treatment of Chagas disease. A series of 26 new compounds waswere designed, synthesized, and tested against the recombinant cruzain (Cz) to map its S1/S1' subsites. The same series was evaluated on a panel of four human cysteine proteases (CatB, CatK, CatL, CatS) and Leishmania mexicana CPB, which is a potential target for the treatment of cutaneous leishmaniasis. The synthesized compounds are dipeptidyl nitriles designed based on the most promising combinations of different moieties in P1 (ten), P2 (six), and P3 (four different building blocks). Eight compounds exhibited a Ki smaller than 20.0 nM for Cz, whereas three compounds met these criteria for LmCPB. The three inhibitors had an EC50 value of ca. 4 M, thus being equipotent to benznidazole according to the anti-trypanosomal effects. Our mapping approach and the respective structure-activity relationships provide insights into the specific ligand-target interactions for therapeutically relevant cysteine proteases.\n\nAuthor SummaryDespite many achievements in identifying novel agents for the treatment of tropical and neglected diseases, further research continues to be of fundamental importance. Our research groups have been using the cruzipain cysteine protease in its recombinant form, cruzain (Cz), to identify new trypanocidal agents. Considering the possible interchangeability with other cysteine proteases, the same series of dipeptidyl nitriles was tested in Leishmania mexicana LmCPB. Other potential targets for such inhibitors are human cysteine cathepsins, which are involved in different disease states. Thus, the inhibitors were also tested against cathepsins B, L, K, and S. Our results demonstrate that inhibition of these cysteine proteases can be achieved by appropriate structural modifications of dipeptidyl nitriles. It was also possible to identify trypanocidal agents, equipotent to benznidazole, the current drug of choice used for the treatment of Chagas disease.

biochemistry↗

bioRxiv2018

Genotyping by sequencing of 393 Sorghum bicolor BTx623 x IS3620C recombinant inbred lines improves sensitivity and resolution of QTL detection

We describe a genetic map with a total of 381 bins of 616 genotyping by sequencing (GBS)-based SNP markers in a F6-F8 recombinant inbred line (RIL) population of 393 individuals derived from crossing S. bicolor BTx623 to S. bicolor IS3620C, a guinea line substantially diverged from BTx623. Five segregation distorted regions were found with four showing enrichment for S. bicolor alleles, suggesting possible selection during formation of this RIL population. A quantitative trait locus (QTL) study with this number of individuals, tripled relative to prior studies of this cross, provided resources, validated previous findings, and demonstrated improved power to detect plant height and flowering time related QTLs relative to other published studies. An unexpected low correlation between flowering time and plant height permitted us to separate QTLs for each trait and provide evidence against pleiotropy. Ten non-random syntenic regions conferring QTLs for the same trait suggest that those QTLs may represent alleles at genes functioning in the same manner since the 96 million year ago genome duplication that created these syntenic relationships, while syntenic regions conferring QTLs for different trait may suggest sub-functionalization after duplication. Collectively, this study provides resources for marker-assisted breeding, as well as a framework for fine mapping and subsequent cloning of major genes for important traits such as plant height and flowering time in sorghum.

genetics↗