Combinatorial Design of a Nanobody that Specifically Targets Structured RNAs

Cawez, Frédéric; Duray, Elodie; Hu, Yaozhong; Vandenameele, Julie; Romão, Ema; Vincke, Cécile; Dumoulin, Mireille; Galleni, Moreno; Vandevenne, Marylène

doi:10.1016/j.jmb.2018.03.032

Cited by 13 publications

(9 citation statements)

References 55 publications

(63 reference statements)

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“…For the construction of the synthetic library, VHH genes were designed in which codons of complementarity determining regions (CDRs) were randomized with a bias for Ser, Tyr, Arg, Lys, Gly, and Asn (Figure 1). About 9.63 × 10 10 individual transformants were obtained for the synthetic library [36]. To ensure the quality of the library, a colony PCR was performed on 100 randomly selected clones, and the result after agarose gel electrophoresis revealed the presence of correctly sized insert for a VHH gene in the phagemids within 80% of the clones.…”

Section: Construction Of the Libraries Panning And Screeningmentioning

confidence: 99%

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Intrabody Targeting HIF-1α Mediates Transcriptional Downregulation of Target Genes Related to Solid Tumors

Romão

Vincke

et al. 2021

IJMS

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Uncontrolled growth of solid tumors will result in a hallmark hypoxic condition, whereby the key transcriptional regulator of hypoxia inducible factor-1α (HIF-1α) will be stabilized to activate the transcription of target genes that are responsible for the metabolism, proliferation, and metastasis of tumor cells. Targeting and inhibiting the transcriptional activity of HIF-1 may provide an interesting strategy for cancer therapy. In the present study, an immune library and a synthetic library were constructed for the phage display selection of Nbs against recombinant PAS B domain protein (rPasB) of HIF-1α. After panning and screening, seven different nanobodies (Nbs) were selected, of which five were confirmed via immunoprecipitation to target the native HIF-1α subunit. The inhibitory effect of the selected Nbs on HIF-1 induced activation of target genes has been evaluated after intracellular expression of these Nbs in HeLa cells. The dramatic inhibition of both intrabody formats on the expression of HIF-1-related target genes has been confirmed, which indicated the inhibitory efficacy of selected Nbs on the transcriptional activity of HIF-1.

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Section: Construction Of the Libraries Panning And Screeningmentioning

confidence: 99%

“…For the construction of the synthetic library, the amino acid sequence was designed based on the scaffold of Nb BCII10, and the sequence of the CDRs was determined based on the sequence analysis of naturally occurring Nbs [36]. Bias was given to certain amino acids (i.e., Lys, Arg, Ser, Tyr, Asn, and Gly).…”

Section: Construction Of Nb Librariesmentioning

confidence: 99%

Intrabody Targeting HIF-1α Mediates Transcriptional Downregulation of Target Genes Related to Solid Tumors

Romão

Vincke

et al. 2021

IJMS

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“…They have both high affinity and specificity and the potential to interrupt lncRNA-RBP interactions [308]. Nanobodies can be designed to specifically target highly structured RNA molecules [309], and since it is known that many lncRNAs such as MALAT1 [310], NEAT1 [311], XIST [230], or HOTAIR [312] are well-structured, this approach could be used for treating OC types in which those lncRNAs are overexpressed. Small molecules (chemical compounds), by binding to either lncRNAs or RNA-binding proteins (RBPs), can change their secondary or tertiary structures or mask protein-binding sites on lncRNAs or lncRNA-binding domains of RBPs, and thus disrupt interactions between them [313,314].…”

Section: Strategies For Targeting Lncrnas As a Treatment For Ocsmentioning

confidence: 99%

(In)Distinctive Role of Long Non-Coding RNAs in Common and Rare Ovarian Cancers

Sabol

Calleja‐Agius

Fiore

et al. 2021

Cancers

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Rare ovarian cancers (ROCs) are OCs with an annual incidence of fewer than 6 cases per 100,000 women. They affect women of all ages, but due to their low incidence and the potential clinical inexperience in management, there can be a delay in diagnosis, leading to a poor prognosis. The underlying causes for these tumors are varied, but generally, the tumors arise due to alterations in gene/protein expression in cellular processes that regulate normal proliferation and its checkpoints. Dysregulation of the cellular processes that lead to cancer includes gene mutations, epimutations, non-coding RNA (ncRNA) regulation, posttranscriptional and posttranslational modifications. Long non-coding RNA (lncRNA) are defined as transcribed RNA molecules, more than 200 nucleotides in length which are not translated into proteins. They regulate gene expression through several mechanisms and therefore add another level of complexity to the regulatory mechanisms affecting tumor development. Since few studies have been performed on ROCs, in this review we summarize the mechanisms of action of lncRNA in OC, with an emphasis on ROCs.

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“…Therefore, nanobodies can be engineered to recognize stRNA epitopes. Nevertheless, their specificity should be improved in future works [104].…”

Section: Nanobodiesmentioning

confidence: 99%

Strategies to target long non-coding RNAs in cancer treatment: progress and challenges

Dizaji

2020

Egypt J Med Hum Genet

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Background: Long non-coding RNAs are important regulators of gene expression and diverse biological processes. Their aberrant expression contributes to a verity of diseases including cancer development and progression, providing them with great potential to be diagnostic and prognostic biomarkers and therapeutic targets. Therefore, they can have a key role in personalized cancer medicine. This review aims at introducing possible strategies to target long ncRNAs therapeutically in cancer. Also, chemical modification of nucleic acid-based therapeutics to improve their pharmacological properties is explained. Then, approaches for the systematic delivery of reagents into the tumor cells or organs are briefly discussed, followed by describing obstacles to the expansion of the therapeutics. Main text: Long ncRNAs function as oncogenes or tumor suppressors, whose activity can modulate all hallmarks of cancer. They are expressed in a very restricted spatial and temporal pattern and can be easily detected in the cells or biological fluids of patients. These properties make them excellent targets for the development of anticancer drugs. Targeting methods aim to attenuate oncogenic lncRNAs or interfere with lncRNA functions to prevent carcinogenesis. Numerous strategies including suppression of oncogenic long ncRNAs, alternation of their epigenetic effects, interfering with their function, restoration of downregulated or lost long ncRNAs, and recruitment of long ncRNAs regulatory elements and expression patterns are recommended for targeting long ncRNAs therapeutically in cancer. These approaches have shown inhibitory effects on malignancy. In this regard, proliferation, migration, and invasion of tumor cells have been inhibited and apoptosis has been induced in different cancer cells in vitro and in vivo. Downregulation of oncogenic long ncRNAs and upregulation of some growth factors (e.g., neurotrophic factor) have been achieved. Conclusions: Targeting long non-coding RNAs therapeutically in cancer and efficient and safe delivery of the reagents have been rarely addressed. Only one clinical trial involving lncRNAs has been reported. Among different technologies, RNAi is the most commonly used and effective tool to target lncRNAs. However, other technologies need to be examined and further research is essential to put lncRNAs into clinical practice.

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Combinatorial Design of a Nanobody that Specifically Targets Structured RNAs

Cited by 13 publications

References 55 publications

Intrabody Targeting HIF-1α Mediates Transcriptional Downregulation of Target Genes Related to Solid Tumors

Intrabody Targeting HIF-1α Mediates Transcriptional Downregulation of Target Genes Related to Solid Tumors

(In)Distinctive Role of Long Non-Coding RNAs in Common and Rare Ovarian Cancers

Strategies to target long non-coding RNAs in cancer treatment: progress and challenges

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