The Needle Issue #13


Welcome to The Needle, a newsletter from Haystack Science to help you navigate the latest translational research, with a roundup of the latest news on preclinical biotech startups from around the world.

In this issue we take a look at a breakthrough in sequence-based design of peptide binders in the context of the stampede of startups developing AI/machine learning-based peptide design platforms guided by structural data. In the translational literature, E3 ubiquitin ligase biology caught our attention, together with several interesting neuroimmunology studies exploring links between the nervous system and cancer progression. Also, several new venture funds for early-stage were announced together with some accelerator/incubator initiatives. Preclinical licensing and partnering have been slow during August, although a sprinkling of financings were announced. Any that we missed, let us know (info@haystacksci.com).

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While most parts of biotech early-stage financing have been in the doldrums in the past two or three years, so-called tech-bio startups have been thriving. Since the posterchild $1.0 billion mega series A round last April of Xaira Therapeutics, which was founded by scientists out of Nobel prize winner David Baker’s group at the University of Washington, several startups seeking to develop machine learning models for designing miniproteins or peptide binders of challenging or ‘undruggable’ targets have emerged, including Enlaza Therapeutics, Vilya, and UbiquiTx. All of these have been developing their own proprietary models based on Alphafold 3, Boltz-1 or Chai-1 for structure prediction and tools based off RFdiffusion, Bindcraft and ProteinMPNN for peptide design. Predicting CDR loops for de novo antibody design is a considerably more challenging task than for simple peptides, but Nabla Bio, founded last year by scientists out of George Church’s lab at Harvard, claims it is doing just that for GPCRs and ion channels. Earlier this month, Chai Discovery also launched with a $100 million series A from Menlo Ventures to optimize multimodal generative models such as Chai-2, which, according to the company, already “achieves a 16% hit rate in de novo antibody design.”

Designing peptides that can selectively bind to a protein target and show therapeutic activity remains a challenge, however, as it often depends on the availability of high-quality structural information about the target molecule, which is seldom available for many disease-relevant proteins that are unstructured or conformationally disordered. Similarly modeling protein-protein interactions like antibody-antigen interactions that are extremely dynamic and floppy also poses problems. All of which raises the question as to whether binders could be predicted simply using amino acid sequence information instead of structural data.

Now, a team led by Pranam Chatterjee from Duke University has addressed this question. In a recent paper in Nature Biotechnology, Chatterjee and his collaborators report the creation of PepMLM, a peptide binder design algorithm based on masked language modeling. A key feature of the algorithm is that it depends exclusively on protein sequence, not structure. Built upon the ESM-2 (Evolutionary Scale Modeling 2) protein language model, PepMLM masks and reconstructs entire peptide regions appended to target protein sequences. This design compels the model to generate context-specific binders. To train PepMLM, the team used high-quality curated datasets from PepNN and Propedia comprising ~10k putative peptide-protein sequence pairs. PepMLM output was consistently found to outperform RFDiffusion on held-out/structured targets, with a higher hit rate (38% to 29%) and low perplexities that closely matched real binders, with generated sequences showing target specificity, even in stringent permutation tests.

The model generated binders predicted to have higher binding scores than native and structure-based binders designed through other methods. Indeed, in vitro validation experiments confirmed the high affinity and specificity of PepMLM-generated binders.

Chatterjee and his colleagues went on to turn their binders into degraders by fusing them to E3 ubiquitin ligase domains, such as CHIP/STUB1. When tested in vitro, over 60% of these degraders knocked down their target proteins. PepMLM peptides achieved nanomolar binding affinity on the drug targets neural cell adhesion molecule 1 (NCAM1), a key marker of acute myeloid leukemia, and anti-Müllerian hormone type 2 receptor (AMHR2), a critical regulator of polycystic ovarian syndrome (where RFDiffusion-predicted peptides failed to bind). The authors also demonstrated that PepMLM-predicted peptides fused to E3 ubiquitin ligases not only degraded MSH3 but completely eliminated mutant huntingtin protein exon 1 containing 43 CAG repeats in Huntington disease patient-derived fibroblast cells. Similar results were obtained for a PepMLM-predicted peptide binder of MESH1, a protein controlling ferroptosis, in collaboration with Ashley Chi Jen-Tsan’s group at Duke University (RFDiffusion again gave no hits). And with Madelaine Dumas and Hector Aguilar-Carreno’s group, in collaboration with Matt Delisa's group at Cornell University, PepMLM-derived peptides bound and reduced levels of viral phosphoproteins from Nipah, Hendra, and human metapneumovirus (HMPV); indeed, in live HMPV infection models, the PepMLM peptide mediated high levels of P protein clearance.

The ability of PepMLM to design binders purely on the basis of target-protein sequence is an important advance towards designing therapeutic peptides against hitherto inaccessible targets that lack structural data. Future work should explore how to incorporate chemical modifications such as cyclization or stapling to enhance stability of the binders, as well as the evaluation of the strongest candidates in vivo. Another challenge will be to ameliorate the immunogenicity of these foreign de novo proteins. The use of protein engineering approaches, such as incorporation of mirror amino acids that can cloak foreign peptides from the immune system, may offer solutions. But it is likely that candidates discovered using sequence or structure prediction tools will still require lengthy development programs to be turned into safe and effective drugs, despite the hype.

Papers: Best of the rest

Target biology

ZBTB11 synthetic lethal dependency in KRAS inhibitor-resistant pancreatic ductal adenocarcinoma targeted in vitro by molecular glues | Nature Chemical Biology

d-cysteine impairs tumour growth in vitro and in breast cancer xenograft models by inhibiting cysteine desulfurase NFS1 | Nature Metabolism

CRISPR/Cas9 knockout of NASP, a stabilizer of histones H3 and H4, in chronic myeloid leukemia/breast cancer cell lines and breast cancer xenograft mouse models overcomes PARP inhibitor resistance | Nature

CGRP-related neuropeptide adrenomedullin 2 promotes tissue-protective type 2 innate lymphoid cell (ILC2) responses and limits intestinal inflammation in mouse colon cell lines and irritible bowel syndrome models | Nature Immunology

E3 ubiquitin ligase Listerin regulates macrophage cholesterol efflux and atherosclerosis progression by catalyzing K63-linked polyubiquitination of membrane protein transporter ABCA1 | JCI

Macrocyclic peptide cyclin A/B RxL inhibitors block G1–S-checkpoint-compromised E2F signaling in mouse models of small cell lung cancer | Nature

CRISPRi, CRISPRa and CRISPRcutting screening together with single-cell transcriptomics in primary human 3D gastric organoids identifies identifies transcription initiator TAF6L as keyregulator of fucosylation and resistance to cisplatin chemotherapy | Nature Communications

Intriguing interactions between the nervous system and cancer progression

Presynaptic acetylcholine signaling through muscarinic receptor CHRM3 input promotes progression in glioblastoma patient-derived xenografts | Cancer Cell

Axonal injury mediated via enzyme SARM1 signaling is a driver of progression in glioblastoma patient-derived xenografts | Nature

Cancer-induced nerve injury promotes resistance to anti-PD-1 therapy in squamous cell carcinoma | Nature

Cancer immunology

Genome-wide CRISPR screens of CD8+ T cells exposed to melanoma cells identify E3 ubiquitin ligase STUB1–adaptor protein CHIC2 complex as targets to boost T-cell mediated anti-tumor immunity | Nature Immunology

Genome-wide CRISPR screens of NK cells multiply exposed to pancreatic ductal adenocarcinoma cells identify mediator complex components MED12 and CCNC or E3 ubiquitin ligase ARIH2 as targets for enhancing CAR-NK cell antitumor potency | Cancer Cell

Chemical library screening and CRISPRi screening inhibition of macropinocytosis in human colorectal adenocarcinoma, pancreatic cancer and mouse breast cancer cells identifies dihydroorotate dehydrogenase (DHODH) as driver of immunosuppression and PD-1 resistance | Immunity

E3 ubiquitin ligase TRIM24 is a therapeutic target in endocrine treatment–resistant breast cancer cell lines and patient-derived organoids | PNAS

Macrophage Tank-binding kinase 1 (TBK1) expression levels correlate poor survival in breast cancer brain metastasis patients and drive development and outgrowth in mouse models | PNAS

Loss of m6A reader protein YTHDF2 enhances T-helper 9 (Th9) programming and CAR-Th9 cell antitumor efficacy in lung adenocarcinoma mouse xenograft models | Nature Immunology

T cell receptor (TCR)-sufficient anti-CD19 allogeneic CAR-T cells with knockout of signal peptide peptidase-like 3 (SPPL3) show altered cell surface glycan and enhanced anti-tumor activity in lymphoma and leukemia patients | Cell

Pharmacological or genetic perturbation of calreticulin potentiates CD8+ T cell–mediated antitumor responses in vitro | Journal of Experimental Medicine

Proof-of concept studies

Targeting the MYC oncogene with a selective bi-steric mTORC1 inhibitor elicits tumor regression in a MYC-driven genetically engineered mouse model and hepatocellular carcinoma patient-derived xenografts cancers | Cell Chemical Biology

Discovery of indole acyl sulfonamide derivatives of β-carboline-1-propionic acid isolated from tropical plant Eurycoma longifolia Jack through phenotypic screening and structural optimization | Nature Communications

BET inhibitors reduce tumor growth in preclinical models of gastrointestinal gene signature–positive castration-resistant prostate cancer | JCI

De novo design of a peptide modulator reverses sodium channel dysfunction in NaV1.5 model of cardiac arrhythmia and SCN8A model of epilepsy | Cell

Targeting the MYC oncogene with a selective bi-steric mTORC1 inhibitor elicits tumor regression in MYC-driven cancers | Cell Chemical Biology

Bezafibrate improves mitochondrial function and blood-brain barrier integrity in patient-derived brain microvascular endothelial cells and improves social deficits in mouse models of 22q11.2 deletion syndrome | Science Translational Medicine

Genome editing

In vivo genome editing of human haematopoietic stem cells for treatment of blood disorders using mRNA delivery | Nature Biomedical Engineering

In vivo base editing delivered via AAV-8 reduces liver cysts in mouse model of autosomal dominant polycystic kidney disease | Molecular Therapy

Engraftment and persistence of HBB base-edited hematopoietic stem cells in nonhuman primates | Science Translational Medicine

CNS-targeted base editing of the major late-onset Tay-Sachs mutation alleviates disease in mice | JCI

Single-dose genome editing therapy rescues auditory and vestibular functions in adult mice with DFNA41 deafness | JCI

Platform technologies

Target-adjacent motif (TAM) interaction domain (TID) deletion converts IscB and Cas9 into RNA-guided RNA editors | Cell

Method for enhancing glycan targeting via structure-guided site-directed mutagenesis and directed evolution of affinity-matured mAb binders of cancer cell membrane polysialic acids tested in a small cell lung cancer cell line | Cell Chemical Biology

Lipid nanoparticles comprising sphingomyelin conjugated to paclitaxel combined with ultra-pH-sensitive azepane and decorated with CD47 ‘do not eat me’ peptide (CD47p) enhance half-life and efficacy of gemcitabine or carboplatin in triple-negative breast cancer and pancreatic cancer mouse models | Nature Cancer

Startup news

​Three more early-stage funds were announced:

Hatteras Venture partners raises >$200 million for two new fund on seed-/early-stage startups in biotech, medtech, and healthtech

Aarhus University launches $94 million Delphinus Venture Capital fund to back spinouts and startups in east and central Denmark, including biotech.

Medical University of South Carolina devotes $30 million to venture funds dedicated to innovative health care startups

Meanwhile, several incubators made announcements of funding or startups:

BioMed X Heidelburg collaboration with Novo Nordisk invites scientists worldwide to submit project proposals for peptide oral delivery by October 12, 2025

University of Cambridge unveils inaugural SPARK cohort, including therapeutics startups SafeTide and Yacson Therapeutcs

65LAB partnership of ClavystBio, Leaps by Bayer, Lightstone Ventures, Polaris Partners/Polaris Innovation Fund and Evotec award $1.5 M to Duke-National University of Singapore to fund work leading to spin out focusing on antifibrotic drugs

Among all the other layoff news, one year after launching “biotech Bell Labs”, Arena Bioworks has had to retrench:

Arena BioWorks lays off 30% of staff to move out of cell and gene therapy

Preclinical funding

Preclinical deals

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Until next week,

Juan Carlos and Andy

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